Nuova ricerca

CARLO PINCELLI

COLLABORATORE DI RICERCA
Dipartimento Chirurgico, Medico, Odontoiatrico e di Scienze Morfologiche con interesse Trapiantologico, Oncologico e di Medicina Rigenerativa

Home |


Pubblicazioni

- Composizioni contenenti nerve growth factor per la prevenzione del danno prodotto dall'esposizione ai raggi ultravioletti [Brevetto]
Pincelli, Carlo
abstract

Si descrivono composizioni cosmetiche e farmaceutiche contenenti NGF per la prevenzione del danno indotto dall'esposizione ai raggi ultravioletti

- Topical use of tyrosine kinase inhibitors of microbial origin to prevent and treat skin disorders characterised by excesssive cell proliferation [Brevetto]
Pincelli, Carlo
abstract

The present invention relates to the use of the alkaloid K252 and its analogues or derivatives to prepare topical drugs for the treatment of disorders characterised by hyperproliferation of keratinocytes

- Uso topico di inibitori della tirosina chinasi di origine microbica per prevenire e combattere le patologie dermatologiche da eccesso di proliferazione cellulare [Brevetto]
Pincelli, Carlo
abstract

La presente invenzione riguarda l'impiego del K252, inibitore delle tirosino chinasi, per la preparazione farmaceutica per uso topico nella terapia delle patologie iperproliferative e della psoriasi

2023 - A Novel In Vivo Active Pemphigus Model Targeting Desmoglein1 and Desmoglein3: A Tool Representing All Pemphigus Variants [Articolo su rivista]
Lotti, Roberta; Atene, Claudio Giacinto; Zanfi, Emma Dorotea; Bertesi, Matteo; Pincelli, Carlo; Zanocco-Marani, Tommaso
abstract

: Background: Pemphigus is a life-threatening blistering autoimmune disease. Several forms, characterized by the presence of autoantibodies against different autoantigens, have been described. In Pemphigus Vulgaris (PV), autoantibodies target the cadherin Desmoglein 3 (DSG3), while in Pemphigus foliaceous (PF) autoantibodies target the cadherin Desmoglein 1 (DSG1). Another variant, mucocutaneous Pemphigus, is characterized by the presence of IgG against both DSG1 and DSG3. Moreover, other forms of Pemphigus characterized by the presence of autoantibodies against other autoantigens have been described. With regard to animal models, one can distinguish between passive models, where pathological IgG are transferred into neonatal mice, and active models, where B cells deriving from animals immunized against a specific autoantigen are transferred into immunodeficient mice that develop the disease. Active models recreate PV and a form of Pemphigus characterized by the presence of IgG against the cadherin Desmocollin 3 (DSC3). Further approaches allow to collect sera or B/T cells from mice immunized against a specific antigen to evaluate the mechanisms underlying the onset of the disease. Objective: To develop and characterize a new active model of Pemphigus where mice express auto antibodies against either DSG1 alone, or DSG1 and DSG3, thereby recapitulating PF and mucocutaneous Pemphigus, respectively. In addition to the existing models, with the active models reported in this work, it will be possible to recapitulate and mimic the main forms of pemphigus in adult mice, thus allowing a better understanding of the disease in the long term, including the benefit/risk ratio of new therapies. Results: The new DSG1 and the DSG1/DSG3 mixed models were developed as proposed. Immunized animals, and subsequently, animals that received splenocytes from the immunized donors produce a high concentration of circulating antibodies against the specific antigens. The severity of the disease was assessed by evaluating the PV score, evidencing that the DSG1/DSG3 mixed model exhibits the most severe symptoms among those analyzed. Alopecia, erosions, and blistering were observed in the skin of DSG1, DSG3 and DSG1/DSG3 models, while lesions in the mucosa were observed only in DSG3 and DSG1/DSG3 animals. The effectiveness of the corticosteroid Methyl-Prednisolone was evaluated in the DSG1 and DSG1/DSG3 models, that showed only partial responsiveness.

2023 - Blocking soluble Fas Ligand ameliorates pemphigus: PC111 efficacy in ex-vivo human pemphigus models [Articolo su rivista]
Lotti, Roberta; Hundt, Jennifer E; Ludwig, Ralf J; Bennett, Brydon; Amato, Antonino; Marconi, Alessandra; Pincelli, Carlo
abstract

: Pemphigus is a life-threatening, chronic, autoimmune bullous disease affecting both the skin and the mucous membranes. Based on the mainstream concept that blister formation occurs upon binding of autoantibodies to their antigen proteins (desmoglein1, DSG1 and desmoglein3, DSG3), current therapies mostly aim to suppress the immune system. To avoid the severe side effects associated with the chronic use of immunosuppressive treatments, we have developed PC111, a fully human monoclonal antibody targeting human Fas ligand (FasL). We have provided a number of in vitro and in vivo evidences showing that soluble FasL induces keratinocyte apoptosis followed by acantholysis. An anti-murine FasL prevents blister formation in the pemphigus neonatal mouse model. To confirm the mechanism of action (MoA) and the efficacy of PC111 in a human pemphigus context, we used the keratinocyte dissociation assay and two independent Human Skin Organ Cultures (HSOC) pemphigus models. PC111 reduced acantholysis in vitro, as shown by the dose-dependent reduction of fragments in the monolayer cultures. In the first HSOC model, normal human skin was subcutaneously injected with a scFv antibody fragment directed against DSG1 and DSG3, resulting in a severe acantholysis (70-100%) after 24 hours. PC111 inhibited blister formation to around 50% of control. In the second model, normal human skin was injected with a mixture of pemphigus patients' autoantibodies resulting in a less severe acantholysis (20-30%). PC111 significantly suppressed blister formation to more than 75% up to 72 hours. These results confirm PC111 MoA and demonstrates the efficacy of the anti-FasL antibody also in a pemphigus setting.

2023 - CD271 activation prevents low to high-risk progression of cutaneous squamous cell carcinoma and improves therapy outcomes [Articolo su rivista]
Quadri, Marika; Tiso, Natascia; Musmeci, Francesco; I Morasso, Maria; R Brooks, Stephen; REGGIANI BONETTI, Luca; Panini, Rossana; Lotti, Roberta; Marconi, Alessandra; Pincelli, Carlo; Palazzo, Elisabetta
abstract

2022 - Isolation of an "early" transit amplifying keratinocyte population in human epidermis: a role for the low affinity neurotrophin receptor CD271 [Articolo su rivista]
Lotti, Roberta; Palazzo, Elisabetta; Quadri, Marika; Dumas, Marc; Schnebert, Sylvianne; Biondini, Diego; Bianchini, Maria Anastasia; Nizard, Carine; Pincelli, Carlo; Marconi, Alessandra
abstract

In the interfollicular epidermis (IFE), stem cells (KSC) generate transit amplifying (TA) cells that, after symmetric divisions, produce differentiating daughters. Here, we isolated and characterized the highly proliferative interfollicular epidermal basal cell population "early" TA (ETA) cells, based on their capacity to adhere to type IV collagen. Proliferation and colony-forming efficiency in ETA cells are lower than in KSC but higher than in "late" TA (LTA). Stemness, proliferation, and differentiation markers confirmed that ETA cells display a unique phenotype. Skin reconstructs derived from ETA cells present different features (epidermal thickness, Ki67, and Survivin expression), as compared to skin equivalents generated from either KSC or LTA cells. The low-affinity neurotrophin receptor CD271, which regulates the KSC to TA cell transition in the human epidermis through an on/off switch control mechanism, is predominantly expressed in ETA cells. Skin equivalents generated from siRNA CD271 ETA cells display a more proliferative and less differentiated phenotype, as compared to mock-derived reconstructs. Consistently, CD271 overexpression in LTA cells generates a more proliferative skin equivalent than mock LTA cells. Finally, the CD271 level declines with cellular senescence, while it induces a delay in p16INK4 expression. We conclude that ETA cells represent the first KSC progenitor with exclusive features. CD271 identifies and modulates ETA cells, thus participating in the early differentiation and regenerative capacity of the human epidermis.

2021 - A Novel Multi-Action Emollient Plus Cream Improves Skin Barrier Function in Patients with Atopic Dermatitis: In vitro and Clinical Evidence [Articolo su rivista]
Quadri, Marika; Lotti, Roberta; Bonzano, Laura; Ciardo, Silvana; Guanti, Mario Bruno; Pellacani, Giovanni; Pincelli, Carlo; Marconi, Alessandra
abstract

Emollients capable of restoring the skin barrier function would extend their role beyond basic maintenance therapy in atopic dermatitis (AD).

2021 - Dermocosmetics: beneficial adjuncts in the treatment of acne vulgaris [Articolo su rivista]
Araviiskaia, E.; Lopez Estebaranz, J. L.; Pincelli, C.
abstract

Introduction: Dermocosmetics are increasingly being recognized as an integral part of acne management. Dermocosmetics may minimize the side effects of acne medications, provide synergistic effects by improving the efficacy of other treatments, and limit exposure to environmental factors such as ultraviolet radiation. We aimed to provide an overview of the active ingredients and different types of preparations used in dermocosmetics for acne, and highlight supporting evidence for their use in clinical practice. Methods: A literature search for selected key words was performed using PubMed. Additional papers were identified based on author expertize. Results and discussion: The different types of active ingredients in dermocosmetics for acne can be classified as: sebum-controlling, antimicrobial, anti-inflammatory, anti-oxidant and/or keratolytic. Such agents may modulate the pathogenic pathways in acne. Dermocosmetics can be formulated as emulsions/creams, cleansers or camouflaging make-up. Dermocosmetics are useful treatment adjuncts for acne and have been shown to improve the clinical signs of acne, reduce transepidermal water loss and modify sebum production. Dermocosmetics have also been associated with reducing side effects of pharmacological treatments, high levels of patient satisfaction and increased adherence to treatment regimens. Together this evidence supports the use of dermocosmetics in clinical practice.

2021 - Promoter Methylation Leads to Decreased ZFP36 Expression and Deregulated NLRP3 Inflammasome Activation in Psoriatic Fibroblasts [Articolo su rivista]
Bertesi, M.; Fantini, S.; Alecci, C.; Lotti, R.; Martello, A.; Parenti, S.; Carretta, C.; Marconi, A.; Grande, A.; Pincelli, C.; Zanocco Marani, T.
abstract

The mRNA-destabilizing protein tristetraprolin (TTP), encoded by the ZFP36 gene, is known to be able to end inflammatory responses by directly targeting and destabilizing mRNAs encoding pro-inflammatory cytokines. We analyzed its role in psoriasis, a disease characterized by chronic inflammation. We observed that TTP is downregulated in fibroblasts deriving from psoriasis patients compared to those deriving from healthy individuals and that psoriatic fibroblasts exhibit abnormal inflammasome activity compared to their physiological counterpart. This phenomenon depends on TTP downregulation. In fact, following restoration, TTP is capable of directly targeting for degradation NLRP3 mRNA, thereby drastically decreasing inflammasome activation. Moreover, we provide evidence that ZFP36 undergoes methylation in psoriasis, by virtue of the presence of long stretches of CpG dinucleotides both in the promoter and the coding region. Besides confirming that a perturbation of TTP expression might underlie the pathogenesis of psoriasis, we suggest that deregulated inflammasome activity might play a role in the disease alongside deregulated cytokine expression.

2021 - Specific activation of the CD271 intracellular domain in combination with chemotherapy or targeted therapy inhibits melanoma progression [Articolo su rivista]
Saltari, Annalisa; Dzung, Andreas; Quadri, Marika; Tiso, Natascia; Facchinello, Nicola; Hernandez-Barranco, Alberto; Garcia-Silva, Susana; Nogués, Laura; Stoffel, Corinne Isabelle; Cheng, Phil F; Turko, Patrick; Eichhoff, Ossia M; Truzzi, Francesca; Marconi, Alessandra; Pincelli, Carlo; Peinado, Héctor; Dummer, Reinhard; Levesque, Mitchell P
abstract

CD271 (NGFR) is a neurotrophin receptor that belongs to the tumor necrosis receptor (TNFR) family. Upon ligand binding, CD271 can mediate either survival or cell death. While the role of CD271 as a marker of tumor-initiating cells is still a matter of debate, its role in melanoma progression has been well documented. Moreover, CD271 has been shown to be upregulated after exposure to both chemotherapy and targeted therapy. In this study, we demonstrate that activation of CD271 by a short β-amyloid-derived peptide (Aβ(25-35)) in combination with either chemotherapy or MAPK inhibitors induces apoptosis in 2D and 3D cultures of 8 melanoma cell lines. This combinatorial treatment significantly reduced metastasis in a zebrafish xenograft model and led to significantly decreased tumor volume in mice. Administration of Aβ(25-35) in ex vivo tumors from immunotherapy- and targeted therapy-resistant patients significantly reduced proliferation of melanoma cells, showing that activation of CD271 can overcome drug resistance. Aβ(25-35) was specific to CD271-expressing cells and induced CD271 cleavage and phosphorylation of JNK (pJNK). The direct protein-protein interaction of pJNK with CD271 led to PARP1 cleavage, p53 and caspase activation, and pJNK-dependent cell death. Aβ(25-35) also mediated mitochondrial reactive oxygen species (mROS) accumulation, which induced CD271 overexpression. Finally, CD271 upregulation inhibited mROS production, revealing the presence of a negative feedback loop in mROS regulation. These results indicate that targeting CD271 can activate cell death pathways to inhibit melanoma progression and potentially overcome resistance to targeted therapy.

2020 - Molecular Approach to Cutaneous Squamous Cell Carcinoma: From Pathways to Therapy [Articolo su rivista]
Palazzo, E; Morasso, Mi; Pincelli, C
abstract

Cutaneous squamous cell carcinoma (cSCC) represents the second most frequent skin cancer,recently showing a rapid increase in incidence worldwide, with around >1 million cases/year in theUnited States and 2500 deaths [1] [...].

2020 - Wnt/CTNNB1 signal transduction pathway inhibits the expression of ZFP36 in squamous cell carcinoma, by inducing transcriptional repressors SNAI1, SLUG and TWIST [Articolo su rivista]
Zanfi, E. D.; Fantini, S.; Lotti, R.; Bertesi, M.; Marconi, A.; Grande, A.; Manfredini, R.; Pincelli, C.; Zanocco-Marani, T.
abstract

The Wnt/CTNNB1 pathway is often deregulated in epithelial tumors. The ZFP36 gene, encoding the mRNA binding protein Tristetraprolin (TTP), is downregulated in several cancers, where it has been described to behave as a tumor suppressor. By this report, we show that Wnt/CTNNB1 pathway is constitutively activated, and ZFP36 expression is downregulated in Squamous Cell Carcinoma (SCC) cell lines compared to normal keratinocytes. Moreover, we suggest that the decrease of ZFP36 expression might depend on the activity of transcriptional repressors SNAI1, SLUG and TWIST, whose expression is induced by Wnt/CTNNB1, highlighting a potential regulatory mechanism underlying ZFP36 downregulation in epithelial cancers.

2019 - Apremilast Normalizes Gene Expression of Inflammatory Mediators in Human Keratinocytes and Reduces Antigen-Induced Atopic Dermatitis in Mice [Articolo su rivista]
Schafer, Peter H; Adams, Mary; Horan, Gerald; Truzzi, Francesca; Marconi, Alessandra; Pincelli, Carlo
abstract

Apremilast, an oral phosphodiesterase (PDE) 4 inhibitor, has demonstrated efficacy in psoriasis, while its efficacy in atopic dermatitis (AD) was found to be modest. AD is a chronic inflammatory skin disease associated with activation of T helper (Th) 2 and Th17 immunity and a compromised epidermal barrier.

2019 - Development of a Desmocollin-3 Active Mouse Model Recapitulating Human Atypical Pemphigus [Articolo su rivista]
Lotti, Roberta; Atene, Claudio Giacinto; Marconi, Alessandra; Di Rocco, Giulia; Reggiani Bonetti, L; Zanocco Marani, Tommaso; Pincelli, Carlo
abstract

Pemphigus vulgaris (PV) is a life-threatening mucocutaneous autoimmune blistering disease. It is often associated with autoantibodies to the desmosomal adhesion proteins Desmoglein 3 (DSG3) and Desmoglein 1 (DSG1). Recently, auto-antigens, such as desmocollins and others have been described in PV and in atypical pemphigus forms such as Pemphigus Herpetiformis (PH), Pemphigus Vegetans (PVeg), and Paraneoplastic Pemphigus (PP). Desmocollins belong to a cadherin subfamily that provides structure to the desmosomes and play an important role in cell-to-cell adhesion. In order to verify the pathogenic activity of anti-Desmocollin 3 (DSC3) antibodies, we developed an active disease model of pemphigus expressing anti-DSC3 autoantibodies or antiDSC3 and anti-DSG3 antibodies. This approach included the adoptive transfer of DSC3 and/or DSG3 lymphocytes to Rag2(-/-) immunodeficient mice that express DSC3 and DSG3. Our results show that the presence of anti-DSC3 auto-antibodies is sufficient to determine the appearance of a pathological phenotype relatable to pemphigus, but with features not completely super-imposable to those observed in the DSG3 active model, suggesting that the DSC3 active model might mimic the atypical pemphigus. Moreover, the presence of both anti-DSC3 and anti-DSG3 antibodies determines a more severe phenotype and a slower response to prednisolone. In conclusion, we have developed an adult DSC3 pemphigus mouse model that differs from the DSG3 model and supports the concept that antigens other than desmogleins may be responsible for different phenotypes in human pemphigus.

2019 - Do DLX3 and CD271 Protect Human Keratinocytes from Squamous Tumor Development? [Articolo su rivista]
Palazzo, Elisabetta; Marconi, Alessandra; Pincelli, Carlo; Morasso, Maria I
abstract

Well-regulated epidermal homeostasis depends on the function of different classes of factors, such as transcription regulators and receptors. Alterations in this homeostatic balance may lead to the development of cutaneous squamous tumorigenesis. The homeobox transcription factor DLX3 is determinant for a p53-dependent regulation of epidermal differentiation and modulates skin carcinogenesis. The maintenance of skin homeostasis also involves the action of neurotrophins (NTs) and their receptors, Trk and CD271. While Trk receptor overexpression is a hallmark of cancer, there are conflicting data on CD271 expression and function in cutaneous SCC (cSCC). Previous studies have reported NT receptors expression in head and neck SSC (HNSCC). We show that CD271 is expressed at low levels in primary cSCC cells and the number of CD271+ cells correlates with cell cohesion in SCC spheroids. In normal epidermis, CD271 is expressed in proliferative progenitor cells and DLX3 in terminally differentiated keratinocytes. Brain-derived neurotrophic factor (BDNF) and neurotrophin 3 (NT3) increase DLX3 expression. In the absence of a functional BDNF receptor TrkB in keratinocytes, we hypothesize that the BDNF-dependent DLX3 response could be mediated via CD271. Altogether, our results support a putative CD271-DLX3 connection in keratinocytes, which might be crucial to preventing squamous skin cancer.

2018 - Mechanisms Causing Loss of Keratinocyte Cohesion in Pemphigus [Articolo su rivista]
Spindler, Volker; Eming, Rüdiger; Schmidt, Enno; Amagai, Masayuki; Grando, Sergei; Jonkman, Marcel F.; Kowalczyk, Andrew P.; Müller, Eliane J.; Payne, Aimee S.; Pincelli, Carlo; Sinha, Animesh A.; Sprecher, Eli; Zillikens, Detlef; Hertl, Michael; Waschke, Jens
abstract

The autoimmune blistering skin disease pemphigus is caused by IgG autoantibodies against desmosomal cadherins, but the precise mechanisms are in part a matter of controversial discussions. This review focuses on the currently existing models of the disease and highlights the relevance of desmoglein-specific versus nondesmoglein autoantibodies, the contribution of nonautoantibody factors, and the mechanisms leading to cell dissociation and blister formation in response to autoantibody binding. As the review brings together the majority of laboratories currently working on pemphigus pathogenesis, it aims to serve as a solid basis for further investigations for the entire field.

2018 - Mechanisms underlying the clinical effects of apremilast for psoriasis [Articolo su rivista]
Pincelli, C.; Schafer, P. H.; French, L. E.; Augustin, M.; Krueger, J. G.
abstract

Psoriasis is a chronic, systemic, inflammatory disease with manifestations resulting from a dysregulated immune response. In psoriatic skin, expression of all phosphodiesterase 4 (PDE4) isoforms (A-D), part of a family of enzymes known to regulate cyclic adenosine monophosphate levels and immune homeostasis, is elevated compared with healthy controls. Agents that inhibit the enzymatic activity of PDE4, the predominant PDE in most immune cells, exert well-recognized anti-inflammatory effects. Apremilast is a selective PDE4 inhibitor approved for the treatment of adults with moderate to severe plaque psoriasis and/or psoriatic arthritis. In vitro and in vivo investigations have demonstrated the beneficial impact of apremilast treatment on PDE4 activity, inflammatory signal expression, and dermal psoriasiform signs. In patients with moderate to severe psoriasis, treatment with apremilast is associated with significant reductions in plasma levels of interleukin (IL)-17F, IL-17A, IL-22, and tumor necrosis factor-α compared with placebo as early as week 4; decreases in cytokine levels were sustained with continued treatment. Multivariate analyses demonstrated that while changes in IL-17F are the most important predictor of improvement in Psoriasis Area and Severity Index scores, apremilast exerts synergistic attenuating effects among a key group of cytokines involved in the pathology of psoriasis, and these effects correlate with improved skin symptoms. These in vitro and clinical data demonstrate that the beneficial effects of apremilast on known inflammatory mediators are associated with its clinical efficacy.

2018 - Progress in melanoma modeling in vitro [Articolo su rivista]
Marconi, Alessandra; Quadri, Marika; Saltari, Annalisa; Pincelli, Carlo
abstract

Melanoma is one of the most studied neoplasia, although laboratory techniques used for investigating this tumor are not fully reliable. Animal models may not predict the human response due to differences in skin physiology and immunity. In addition, international guidelines recommend to develop processes that contribute to the reduction, refinement and replacement of animals for experiments (3Rs). Adherent cell culture has been widely used for the study of melanoma to obtain important information regarding melanoma biology. Nonetheless, these cells grow in adhesion on the culture substrate which differs considerably from the situation in vivo. Melanoma grows in a 3D spatial conformation where cells are subjected to a heterogeneous exposure to oxygen and nutrient. In addition, cell-cell and cell-matrix interaction play a crucial role in the pathobiology of the tumor as well as in the response to therapeutic agents. To better study melanoma new techniques, including spherical models, tumorospheres, and melanoma skin equivalents have been developed. These 3D models allow to study tumors in a microenvironment that is more close to the in vivo situation, and are less expensive and time consuming than animal studies. This review will also describe the new technologies applied to skin reconstructs such as organ-on-a-chip that allows skin perfusion through microfluidic platforms. 3D in vitro models, based on the new technologies, are becoming more sophisticated, representing at a great extent the in vivo situation, the "perfect" model that will allow less involvement of animals up to their complete replacement, is still far from being achieved. This article is protected by copyright. All rights reserved.

2018 - Soluble Fas Ligand is essential for blister formation in Pemphigus [Articolo su rivista]
Lotti, Roberta; Shu, En; Petrachi, Tiziana; Marconi, Alessandra; Palazzo, Elisabetta; Quadri, Marika; Lin, Ann; O’Reillyan, Lorraine A.; Pincelli, Carlo
abstract

Pemphigus is a blistering disease characterized by pemphigus autoantibodies (PVIgG) directed mostly against desmogleins (Dsgs), resulting in the loss of keratinocyte adhesion (acantholysis). Yet, the mechanisms underlying blister formation remain to be clarified. We have shown previously that anti-Fas ligand (FasL) antibody (Ab) prevents PVIgG-induced caspase-8 activation and Dsg cleavage in human keratinocytes, and that sera from pemphigus patients contain abnormally increased levels of FasL. Here, we demonstrate that recombinant FasL induces the activation of caspases prior to Dsg degradation, and anti-FasL Ab prevents acantholysis in cultured keratinocytes. Moreover, the silencing of FasL reduces PVIgG-induced caspase-8 activation and Dsg3 cleavage. Following injection of PVIgG into mice, FasL is upregulated at 1–3 h and is followed by caspase-8-mediated keratinocyte apoptosis, before blister formation. The administration of anti-FasL Ab after PVIgG injection blocks blister formation in mice. Furthermore, we injected PVIgG into two different gene-targeted mutant mice that selectively lack either secreted soluble FasL (sFasL), FasLΔs/Δs mice, or the membrane-bound form of FasL (mFasL), FasLΔm/Δm mice. After PVIgG treatment, blisters are only visible in FasLΔm/Δm animals, lacking mFasL, but still producing sFasL, similar to wild-type (C57BL/6) animals. By contrast, a significant decrease in the relative acantholytic area is observed in the FasLΔs/Δs animals. These results demonstrate that soluble FasL plays a crucial role in the mechanisms of blister formation, and blockade of FasL could be an effective therapeutic approach for pemphigus.

2018 - The 3R approach to experimental dermatology [Articolo su rivista]
Schneider, M. R.; Pincelli, C.
abstract

2017 - Zebrafish as a model organism for the development of drugs for skin cancer [Articolo su rivista]
Bootorabi, F.; Manouchehri, H.; Changizi, R.; Barker, H.; Palazzo, E.; Saltari, A.; Parikka, M.; Pincelli, C.; Aspatwar, A.
abstract

Skin cancer, which includes melanoma and squamous cell carcinoma, represents the most common type of cutaneous malignancy worldwide, and its incidence is expected to rise in the near future. This condition derives from acquired genetic dysregulation of signaling pathways involved in the proliferation and apoptosis of skin cells. The development of animal models has allowed a better understanding of these pathomechanisms, with the possibility of carrying out toxicological screening and drug development. In particular, the zebrafish (Danio rerio) has been established as one of the most important model organisms for cancer research. This model is particularly suitable for live cell imaging and high-throughput drug screening in a large-scale fashion. Thanks to the recent advances in genome editing, such as the clustered regularly-interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) methodologies, the mechanisms associated with cancer development and progression, as well as drug resistance can be investigated and comprehended. With these unique tools, the zebrafish represents a powerful platform for skin cancer research in the development of target therapies. Here, we will review the advantages of using the zebrafish model for drug discovery and toxicological and phenotypical screening. We will focus in detail on the most recent progress in the field of zebrafish model generation for the study of melanoma and squamous cell carcinoma (SCC), including cancer cell injection and transgenic animal development. Moreover, we will report the latest compounds and small molecules under investigation in melanoma zebrafish models.

2017 - p75 Neurotrophin Receptor in the Skin: Beyond Its Neurotrophic Function [Articolo su rivista]
Pincelli, Carlo
abstract

p75 neurotrophin receptor (p75NTR), also known as CD271, is the low-affinity receptor that, together with the tyrosine kinase receptor tropomyosin-receptor kinase (Trk), mediate neurotrophin (NT) functions. Beside their classic role in skin innervation, NT and their receptors constitute a complex cutaneous network associated with a number of autocrine and paracrine activities. In this context, the role of p75NTR is becoming more and more important. This review will focus on the intriguing functions of p75NTR in healthy and diseased skin. First, p75NTR counterbalances the proliferative and survival activities of its cognate receptor Trk by inducing keratinocyte apoptosis. In addition, p75NTR identifies an early transit-amplifying (TA) keratinocyte population and plays a critical role in keratinocyte stem cell transition to its progeny as well as in epidermal differentiation. p75NTR is absent in psoriatic TA cells, thus rendering these cells resistant to apoptosis. On the other hand, p75NTR infection restores NT-induced apoptosis in psoriatic keratinocytes. Taken together, these results provide evidence for a critical role of p75NTR in epidermal homeostasis, while its lack may account for the TA defect in psoriasis. While the issue of p75NTR as a marker of melanoma initiating cells is still to be solved, there is strong evidence that downregulation of this receptor is a precondition to melanoma invasion and metastasis in vitro and in vivo. All in all, this review points to p75NTR as a major actor in both physiologic and pathologic conditions at the skin level.

2016 - CD271 Down-Regulation Promotes Melanoma Progression and Invasion in Three-Dimensional Models and in Zebrafish [Articolo su rivista]
Saltari, Annalisa; Truzzi, Francesca; Quadri, Marika; Lotti, Roberta; Palazzo, Elisabetta; Grisendi, Giulia; Tiso, Natascia; Marconi, Alessandra; Pincelli, Carlo
abstract

CD271 is a neurotrophin receptor variably expressed in melanoma. Although contradictory data are reported on its role as a marker of tumor-initiating cells, little is known about its function in tumor progression. CD271 expression was higher in spheroids derived from freshly isolated cells of primary melanomas and in primary WM115 and WM793-B cell lines, and it decreased during progression to advanced stages in cells isolated from metastatic melanomas and in metastatic WM266-4 and 1205Lu cell lines. Moreover, CD271 was scarcely detected in the highly invasive spheroids (SKMEL28 and 1205Lu). CD271, originally expressed in the epidermis of skin reconstructs, disappeared when melanoma started to invade the dermis. SKMEL8 CD271(-) cells showed greater proliferation and invasiveness in vitro and were associated with a higher number of metastases in zebrafish compared with CD271(+) cells. CD271 silencing in WM115 induced a more aggressive phenotype in vitro and in vivo. On the contrary, CD271 overexpression in SKMEL28 cells reduced invasion in vitro, and CD271 overexpressing 1205Lu cells was associated with a lower percentage of metastases in zebrafish. A reduced cell-cell adhesion was also observed in the absence of CD271. Taken together, these results indicate that CD271 loss is critical for melanoma progression and metastasis.

2016 - Electrochemotherapy induces apoptotic death in melanoma metastases: A histologic and immunohistochemical investigation [Articolo su rivista]
Bigi, Laura; Galdo, Giovanna; Cesinaro, Anna Maria; Vaschieri, Cristina; Marconi, Alessandra; Pincelli, Carlo; Fantini, Fabrizio
abstract

Background: Electrochemotherapy (ECT) is increasingly used in the treatment of primary and secondary skin tumors, but little is known about the pathologic mechanism responsible for tumor cell destruction in humans. Knowledge of detailed mechanism of host response after ECT may improve the treatment efficacy related to patient selection and technique refinements. Aim: The aim of the study was to investigate the histopathology and mechanism of cell death after ECT in cutaneous melanoma metastases. Methods: Skin biopsy specimens were sequentially obtained after ECT of cutaneous melanoma metastases, during a follow-up period of 2 months. Results from histologic evaluation and immunohistochemical characterization of the inflammatory infiltrate (CD3, CD4, CD8, CD56, Granzyme-B) were compared with a panel of apoptosis-related markers. Main outcome measures: Evidence of the mechanism of tumor cell damage, identification of histological and immunohistochemical signs of apoptosis and/or necrosis underlining a possible time course of tumor destruction and inflammatory reaction after ECT. Results: Early signs of epidermal degeneration, an increase of the inflammatory infiltrate, and initial tumor cell morphological changes were already detected 10 min after ECT. The cell damage progression, as demonstrated by histological and immunohistochemical evidence using apoptotic markers (TUNEL and caspase-3 staining), reached a climax 3 days after treatment, to continue until 10 days after. Scarring fibrosis and complete absence of tumor cells were observed in the late biopsy specimens. A rich inflammatory infiltrate with a prevalence of T-cytotoxic CD3/CD8-positive cells was detected 3 h after ECT and was still appreciable 3 months later. Conclusion: This study attempts to define the time course and characteristics of tumor response to ECT. The observations suggest both a direct necrotic cell damage and a rapid activation of apoptotic mechanisms that occur in the early phases of the cutaneous reaction to ECT. A persistent immune response of T-cytotoxic lymphocytes could possibly explain the long-term local tumor control.

2016 - Monopathogenic vs multipathogenic explanations of pemphigus pathophysiology [Articolo su rivista]
Ahmed, A. Razzaque; Carrozzo, Marco; Caux, Frã©dã©ric; Cirillo, Nicola; Dmochowski, Marian; Alonso, Agustã­n Espaã±a; Gniadecki, Robert; Hertl, Michael; Lã³pez-zabalza, Maria J.; Lotti, Roberta; Pincelli, Carlo; Pittelkow, Mark; Schmidt, Enno; Sinha, Animesh A.; Sprecher, Eli; Grando, Sergei A.
abstract

This viewpoint highlights major, partly controversial concepts about the pathogenesis of pemphigus. The monopathogenic theory explains intra-epidermal blistering through the “desmoglein (Dsg) compensation” hypothesis, according to which an antibody-dependent disabling of Dsg 1-and/ or Dsg 3-mediated cell-cell attachments of keratinocytes (KCs) is sufficient to disrupt epidermal integrity and cause blistering. The multipathogenic theory explains intra-epidermal blistering through the “multiple hit” hypothesis stating that a simultaneous and synchronized inactivation of the physiological mechanisms regulating and/or mediating intercellular adhesion of KCs is necessary to disrupt epidermal integrity. The major premise for a multipathogenic theory is that a single type of autoantibody induces only reversible changes, so that affected KCs can recover due to a self-repair. The damage, however, becomes irreversible when the salvage pathway and/or other cell functions are altered by a partnering autoantibody and/or other pathogenic factors. Future studies are needed to (i) corroborate these findings, (ii) characterize in detail patient populations with non-Dsgspecific autoantibodies, and (iii) determine the extent of the contribution of non-Dsg antibodies in disease pathophysiology.

2016 - Phosphodiesterase 4 in inflammatory diseases: Effects of apremilast in psoriatic blood and in dermal myofibroblasts through the PDE4/CD271 complex [Articolo su rivista]
Schafer, Peter H; Truzzi, Francesca; Parton, Anastasia; Wu, Lei; Kosek, Jolanta; Zhang, Ling Hua; Horan, Gerald; Saltari, Annalisa; Quadri, Marika; Lotti, Roberta; Marconi, Alessandra; Pincelli, Carlo
abstract

Phosphodiesterases 4 (PDE4) act as proinflammatory enzymes via degradation of cAMP, whereas PDE4 inhibitors play an anti-inflammatory role in vitro and in vivo. In particular, apremilast has been recently approved for the treatment of psoriasis and psoriatic arthritis. However, little is known on the expression pattern of PDE4 in psoriasis. We report that PDE4B and PDE4D mRNA are overexpressed in peripheral blood mononuclear cells (PBMC) from psoriasis, as compared with normal controls, while apremilast reduces PBMC production of a number of pro-inflammatory cytokines and increases the levels of anti-inflammatory mediators. PDE4 expression is up-regulated in psoriatic dermis as compared with normal skin, with particular regard to fibroblasts. This is confirmed in vitro, where both dermal fibroblasts (DF) and, to a greater extent, myofibroblasts (DM) express all PDE4 isoforms at the mRNA and protein level. Because PDE4 interacts with the nerve growth factor (NGF) receptor CD271 in lung fibroblasts, we evaluated the relationship and function of PDE4 and CD271 in normal human skin fibroblasts. All PDE4 isoforms co-immunoprecipitate with CD271 in DM, while apremilast inhibits apoptosis induced by β-amyloid, a CD271 ligand, in DM. Furthermore, apremilast significantly reduces NGF- and transforming growth factor-β1 (TGF-β1)-induced fibroblast migration, and inhibits DF differentiation into DM mediated by NGF or TGF-β1. Finally, in DM, apremilast significantly reduces cAMP degradation induced by treatment with β-amyloid. Taken together, these results indicate that PDE4 play an important role in psoriasis. In addition, the study reveals that the PDE4/CD271 complex could be important in modulating fibroblast functions.

2016 - Survivin modulates squamous cell carcinoma-derived stem-like cell proliferation, viability and tumor formation in vivo [Articolo su rivista]
Lotti, Roberta; Palazzo, Elisabetta; Petrachi, Tiziana; Dallaglio, Katiuscia; Saltari, Annalisa; Truzzi, Francesca; Quadri, Marika; Puviani, Mario; Maiorana, Antonino; Marconi, Alessandra; Pincelli, Carlo
abstract

Squamous Cell Carcinoma-derived Stem-like Cells (SCC-SC) originate from alterations in keratinocyte stem cells (KSC) gene expression and sustain tumor development, invasion and recurrence. Since survivin, a KSC marker, is highly expressed in SCC-SC, we evaluate its role in SCC-SC cell growth and SCC models. Survivin silencing by siRNA decreases clonal growth of SCC keratinocytes and viability of total, rapidly adhering (RAD) and non-RAD (NRAD) cells from primary SCC. Similarly, survivin silencing reduces the expression of stem cell markers (OCT4, NOTCH1, CD133, β1-integrin), while it increases the level of differentiation markers (K10, involucrin). Moreover, survivin silencing improves the malignant phenotype of SCC 3D-reconstruct, as demonstrated by reduced epidermal thickness, lower Ki-67 positive cell number, and decreased expression of MMP9 and psoriasin. Furthermore, survivin depletion by siRNA in RasG12V-IκBα-derived tumors leads to smaller tumor formation characterized by lower mitotic index and reduced expression of the tumor-associated marker HIF1α, VEGF and CD51. Therefore, our results indicate survivin as a key gene in regulating SCC cancer stem cell formation and cSCC development.

2015 - Apoptosis and Signaling in Bullous Diseases: Pemphigus [Capitolo/Saggio]
Rubenstein, D. S.; Pincelli, C.
abstract

Autoimmune bullous diseases comprise a heterogeneous group of disorders, which not only differ with regard to their immunological characteristics but also with regard to genetic and environmental predisposing factors. A strong association with HLA alleles was found in pemphigus, mucous membrane pemphigoid, pemphigoid gestationis and epidermolysis bullosa acquisita, whereas in bullous pemphigoid, this association appeared much less prominent. Recently, the first non-HLA genes have been associated with pemphigus vulgaris. In pemphigus vulgaris and foliaceus, the potential impact drugs, smoking and vaccination have been described, while in the endemic forms of pemphigus foliaceus, the role of environmental factors, including infectious agents, is emerging.

2015 - CD271 Mediates Stem Cells to Early Progeny Transition in Human Epidermis [Articolo su rivista]
Truzzi, Francesca; Saltari, Annalisa; Palazzo, Elisabetta; Lotti, Roberta; Petrachi, Tiziana; Dallaglio, Katiuscia; Gemelli, Claudia; Grisendi, Giulia; Dominici, Massimo; Pincelli, Carlo; Marconi, Alessandra
abstract

CD271 is the low-affinity neurotrophin (p75NTR) receptor that belongs to the tumor necrosis factor receptor superfamily. Because in human epidermis, CD271 is predominantly expressed in transit-amplifying (TA) cells, we evaluated the role of this receptor in keratinocyte differentiation and in the transition from keratinocyte stem cells (KSCs) to progeny. Calcium induced an upregulation of CD271 in subconfluent keratinocytes, which was prevented by CD271 small interfering RNA. Furthermore, CD271 overexpression provoked the switch of KSCs to TA cells, whereas silencing CD271 induced TA cells to revert to a KSC phenotype, as shown by the expression of β1-integrin and by the increased clonogenic ability. CD271(+) keratinocytes sorted from freshly isolated TA cells expressed more survivin and keratin 15 (K15) compared with CD271(-) cells and displayed a higher proliferative capacity. Early differentiation markers and K15 were more expressed in the skin equivalent generated from CD271(+) TA than from those derived from CD271(-) TA cells. By contrast, late differentiation markers were more expressed in skin equivalents from CD271(-) than in reconstructs from CD271(+) TA cells. Finally, skin equivalents originated from CD271(-) TA cells displayed a psoriatic phenotype. These results indicate that CD271 is critical for keratinocyte differentiation and regulates the transition from KSCs to TA cells.

2015 - Hypoxia-Inducible Factor-1α and CD271 inversely correlate with melanoma invasiveness [Articolo su rivista]
Marconi, Alessandra; Borroni, Riccardo Giovanni; Truzzi, Francesca; Longo, Caterina; Pistoni, Francesca; Pellacani, Giovanni; Pincelli, Carlo
abstract

Melanoma is characterized, among other features, by microenvironmental factors and by an altered apoptotic machinery. Melanoma cell response to a hypoxic environment is transcriptionally regulated by the Hypoxia-Inducible Factor (HIF)-1α. p75 neurotrophin receptor (p75(NTR) ), also called CD271, mediates apoptosis in several cell systems. The purpose of this study was to analyze the expression of HIF-1α and CD271 in melanomas at different phases of progression, as evaluated by histology and reflectance confocal microscopy (RCM). By RCM, 41.67% tumors were characterized by the presence of a population of dendritic and pleomorphic cells (D+P), corresponding to in situ melanoma; 25% exhibited a predominantly round-cell (RN) proliferation with histologic features of superficial melanoma, and 33.33% showed the presence of cells aggregated in nests (DN), typical of invasive melanoma. HIF-1α was scarcely detected in D+P and in RN melanomas, while it was highly expressed in DN tumors. By contrast, CD271 positive cells were mostly detected in D+P population, and barely observed in the other subtypes. This work demonstrates that CD271 expression inversely correlates with hypoxia in melanoma, and that the two markers may be used in the future as diagnostic/prognostic tools for this neoplasm.

2015 - Notch cooperates with survivin to maintain stemness and to stimulate proliferation in human keratinocytes during ageing [Articolo su rivista]
Palazzo, Elisabetta; Morandi, Paolo; Lotti, Roberta; Saltari, Annalisa; Truzzi, Francesca; Schnebert, Sylvianne; Dumas, Marc; Marconi, Alessandra; Pincelli, Carlo
abstract

The Notch signaling pathway orchestrates cell fate by either inducing cell differentiation or maintaining cells in an undifferentiated state. This study aims to evaluate Notch expression and function in normal human keratinocytes. Notch1 is expressed in all epidermal layers, though to a different degree of intensity, with a dramatic decrease during ageing. Notch1 intracellular domain (N1ICD) levels are decreased during transit from keratinocyte stem cells (KSC) to transit amplifying (TA) cells, mimicking survivin expression in samples from donors of all ages. Calcium markedly reduces N1ICD levels in keratinocytes. N1ICD overexpression induces the up-regulation of survivin and the down-regulation of keratin 10 and involucrin, while increasing the S phase of the cell cycle. On the other hand, Notch1 inhibition (DAPT) dose-dependently decreases survivin, stimulates differentiation, and reduces keratinocyte proliferation in samples from donors of all ages. Silencing Notch downgrades survivin and increases keratin 10. In addition, Notch1 inhibition decreases survivin levels and proliferation both in KSC and TA cells. Finally, while survivin overexpression decreases keratinocyte differentiation and increases N1ICD expression both in KSC and TA cells, silencing survivin results in N1ICD down-regulation and an increase in differentiation markers. These results suggest that the Notch1/survivin crosstalk contributes to the maintenance of stemness in human keratinocytes

2015 - Organ culture and Reflectance Confocal Microscopy as new integrated tools for barrier rescue studies in inflammatory skin diseases [Articolo su rivista]
Petrachi, Tiziana; Lotti, Roberta; Palazzo, Elisabetta; Truzzi, Francesca; Saltari, Annalisa; Morandi, Paolo; Ciardo, Silvana; Pellacani, Giovanni; Pincelli, Carlo; Marconi, Alessandra
abstract

Here we present a new integrated approach to understand skin barrier recovery after physical (tape stripping, TS) or chemical (SDS) injury by combining human skin organ culture and Reflectance Confocal Microscopy (RCM). TS in vitro produced a complete removal of stratum corneum and lipids, a drastic decrease of structural and adhesion proteins, and an increase in cell proliferation. Epidermal recovery with either proliferation or differentiation rescue was observed after 18 hours, with no apoptotic cell detection. On the other hand, when skin organ cultures were exposed to 2% SDS, cellular junctions were disrupted and the expression of late differentiation markers decreased. Junctions repair was detected 24 hours after treatment, with the restoration of epidermal integrity. Both models (TP or SDS) showed the induction of immune-inflammatory markers, such as psoriasin, keratin 16, and the increase in Langerhans cell number. RCM confirmed all the morphological and structural features presented by the organ cultures, thus making this technique fast and easily applicable in the context of dermatological research. These results indicate that combination of skin organ models and RCM can be successfully used for the study of barrier perturbation in skin diseases, for toxicology tests, and for evaluating novel therapies.

2014 - Expression of nuclear survivin in normal skin and squamous cell carcinoma: a possible role in tumor invasion [Articolo su rivista]
Dallaglio, Katiuscia; Petrachi, Tiziana; Marconi, Alessandra; Truzzi, Francesca; Lotti, Roberta; Saltari, Annalisa; Morandi, Paolo; M., Puviani; Maiorana, Antonino; Pincelli, Carlo
abstract

Background: Survivin is detected in few adult normal cells and it is highly expressed in cancer. Nuclear survivin facilitates cell cycle entry, while the mitochondrial pool protects cells from apoptosis. Survivin is overexpressed in keratinocyte stem cells (KSC) and protects them from apoptosis. Methods: As KSC are at the origin of squamous cell carcinoma (SCC), we evaluated survivin expression in normal and cancerous skin in vivo by immunohistochemistry and western blotting. HaCaT cells overexpressing survivin and wound-healing assay are used. Anova and Student-T tests are used for statistical analysis. Results: Survivin is localized both in the cytoplasm and in the nucleus of normal adult and young keratinocytes. Nuclear survivin is detected in one every 10/11 basal keratinocytes. When present in suprabasal cells, nuclear survivin is co-expressed with K10, but not with K15 or p75-neurotrophin-receptor (p75NTR), a transit amplifying cell marker. Nuclear, but not cytoplasmic survivin expression dramatically increases in actinic keratosis and in SCC in situ, as compared to normal epidermis, and it is highest in poorly differentiated SCC. In SCC tumors, nuclear survivin-positive cells are mainly K10/p75NTR-negative and K15-positive. In poorly differentiated tumors, survivin mostly localizes in the deep infiltrating areas. When overexpressed in keratinocytes, survivin increases cell migration. Conclusion: High survivin expression and the subcellular localization of survivin correlate with keratinocyte differentiation and are associated with undifferentiated and more invasive SCC phenotype.

2014 - New modulated genes in psoriasis-derived keratinocyte subpopulations. [Abstract in Rivista]
Lotti, Roberta; Tenedini, Elena; Fabiano, A; Truzzi, Francesca; Saltari, Annalisa; Morandi, Paolo; Marconi, Alessandra; Pincelli, Carlo
abstract

T cells play a crucial role in the pathogenesis of psoriasis, recent data emphasize the key role of keratinocytes that, by carrying intrinsic alterations, could determine the formation of skin lesions resembling psoriasis, without the participation of T-cell derived cytokines. In particular, transit amplifying (TA) cells, the stem cell progeny, seems to be responsible for the psoriatic phenotype. The aim of this study was to analyze the role of keratinocytes sub-populations in the pathogenesis of psoriasis. We analyzed the gene expression profile (GEP) of human keratinocyte sub-populations (stem, “early” TA (ETA) and “late” TA (LTA) cells), derived from healthy skin, lesional and non-lesional psoriatic skin. The total RNA samples, extracted from keratinocyte subpopulations immediately after separation, were hybridized onto the Affymetrix human U133 plus 2.0 GeneChip Array. We identified a small number of up-regulated genes (12 probe sets, corresponding to 8 genes) in keratinocyte subpopulations derived from lesional psoriasis vs. healthy and non-lesional psoriasis. We confirmed the increased expression levels of TCN1, S100A7A, KYNU, SERPINB13, FOXE1, but solely due to the keratinocyte component. We identified for the first time the up-regulation of TMEM171, CLEC7A and NDRG4, which seems to correlate with the pathophysiology of psoriasis. Moreover, GEP analysis of lesional psoriasis sub-populations, as compared to the non-lesional psoriatic counterpart revealed the modulation of 17 probe sets, corresponding to 13 genes. Among these genes, we recognized for the first time the up-regulation of IL13RA1, CCDC109B and CD47. These results indicate the importance of keratinocyte compartment in psoriasis, opening the way to the study of new genes potentially critical in the pathogenesis of psoriasis.

2013 - E-FABP induces differentiation in normal human keratinocytes and modulates the differentiation process in psoriatic keratinocytes in vitro. [Articolo su rivista]
Dallaglio, Katiuscia; Marconi, Alessandra; Truzzi, Francesca; Lotti, Roberta; Palazzo, Elisabetta; Petrachi, Tiziana; Saltari, Annalisa; Coppini, Maurizio; Pincelli, Carlo
abstract

Epidermal fatty acid-binding protein (E-FABP) is a lipid carrier, originally discovered in human epidermis. We show that E-FABP is almost exclusively expressed in postmitotic (PM) keratinocytes, corresponding to its localization in the highest suprabasal layers, while it is barely expressed in keratinocyte stem cells (KSC) and transit amplifying (TA) keratinocytes. Transfection of normal human keratinocytes with recombinant (r) E-FABP induces overexpression of K10 and involucrin. On the other hand, E-FABP inhibition by siRNA downregulates K10 and involucrin expression in normal keratinocytes through NF-κB and JNK signalling pathways. E-FABP is highly expressed in psoriatic epidermis, and it is mainly localized in stratum spinosum. Psoriatic PM keratinocytes overexpress E-FABP as compared to the same population in normal epidermis. E-FABP inhibition in psoriatic keratinocytes markedly reduces differentiation, while it upregulates psoriatic markers such as survivin and K16. However, under high-calcium conditions, E-FABP silencing downregulates K10 and involucrin, while survivin and K16 expression is completely abolished. These data strongly indicate that E-FABP plays an important role in keratinocyte differentiation. Moreover, E-FABP modulates differentiation in psoriatic keratinocytes.

2013 - Isolation and Characterization of Squamous Cell Carcinoma-Derived Stem-like Cells: Role in Tumor Formation [Articolo su rivista]
Dallaglio, Katiuscia; Petrachi, Tiziana; Marconi, Alessandra; Truzzi, Francesca; Lotti, Roberta; Saltari, Annalisa; Morandi, Paolo; M., Puviani; Maiorana, Antonino; D. R., Roop; Pincelli, Carlo
abstract

In human epidermis, keratinocyte stem cells (KSC) are characterized by high levels of β1-integrin, resulting in the rapid adhesion to type IV collagen. Since epithelial tumors originate from KSC, we evaluated the features of rapidly adhering (RAD) keratinocytes derived from primary human squamous cell carcinoma of the skin (cSCC). RAD cells expressed higher levels of survivin, a KSC marker, as compared to non-rapidly adhering (NRAD) cells. Moreover, RAD cells proliferated to a greater extent and were more efficient in forming colonies than NRAD cells. RAD cells also migrated significantly better than NRAD cells. When seeded in a silicone chamber and grafted onto the back skin of NOD SCID mice, RAD cells formed tumors 2–4 fold bigger than those derived from NRAD cells. In tumors derived from RAD cells, the mitotic index was significantly higher than in those derived from NRAD cells, while Ki-67 and survivin expression were more pronounced in RAD tumors. This study suggests that SCC RAD stem cells play a critical role in the formation and development of epithelial tumors.

2013 - Keratinocyte Stem Cells: Biology and Clinical Applications [Capitolo/Saggio]
Pincelli, Carlo; Marconi, Alessandra
abstract

Human epidermis represents a large reservoir of stem cells that continue to self-renew throughout life. Stem cells are essential for skin regeneration and for repair after wounding. They allow long-term culture of keratinocytes that produce large sheets of epidermis to cover extensive burns, thus being lifesaving for these patients. Furthermore, stem cells can be expanded in culture, genetically modified to correct the gene deficiency in genetic skin diseases. This chapter will describe the most recent data on stem cell biology and the potential medical applications of these cells.

2013 - Recapitulating atopic dermatitis in three dimensions: cross talk between keratinocytes and nerve fibers [Articolo su rivista]
Pincelli, Carlo; Steinhoff, Martin
abstract

Roggenkamp et al. (2013) report a significant advance in the in vitro reconstruction of atopic dermatitis (AD) by coculturing lesional human keratinocytes with sensory nerve fibers. This work has important implications for understanding the interaction between the resident skin cells and the peripheral nervous system in AD, and it sheds light on the pathways leading to the pruritus that typifies this disease.

2013 - Self-inactivating MLV vectors have a reduced genotoxic profile in human epidermal keratinocytes [Articolo su rivista]
Cavazza, A.; Cocchiarella, F.; Bartholomae, C.; Schmidt, M.; Pincelli, C.; Larcher, F.; Mavilio, F.
abstract

Transplantation of epithelia derived from keratinocyte stem cells transduced by retroviral vectors is a potential therapy for epidermolysis bullosa (EB), a family of inherited skin adhesion defects. The biosafety characteristics of retroviral vectors in keratinocytes are, however, poorly defined. We developed self-inactivating (SIN) vectors derived from the Moloney murine leukemia (MLV) and the human immunodeficiency (HIV) viruses expressing therapeutic levels of LAMB3, a transgene defective in junctional EB, and tested their integration profile in human primary keratinocytes. The SIN-HIV vector showed the expected preference for transcribed genes while the SIN-MLV vector integrated preferentially in regulatory elements, but showed a significantly lower tendency to target cell growth-related genes, transcription start sites and epigenetically defined promoters compared with a wild-type MLV vector in an epithelial cell context. A quantitative gene expression assay in individual keratinocyte clones showed that MLV-derived vectors deregulate expression of targeted genes at a lower frequency than in hematopoietic cells, and that the SIN-MLV design has the lowest activity compared to both MLV and SIN-HIV vectors. This study indicates that SIN-MLV vectors may have a better safety profile in keratinocyte than in hematopoietic cells, and be a reasonable alternative to lentiviral vectors for gene therapy of inherited skin disorders. © 2013 Macmillan Publishers Limited All rights reserved.

2013 - Targeted Gene Addition in Human Epithelial Stem Cells by Zinc-finger Nuclease-mediated Homologous Recombination [Articolo su rivista]
Coluccio, Andrea; Miselli, Francesca; Angelo, Lombardo; Marconi, Alessandra; Guidantonio Malagoli, Tagliazucchi; Manuel A., Gonçalves; Pincelli, Carlo; Giulietta, Maruggi; Marcela Del, Rio; Luigi, Naldini; Fernando, Larcher; Mavilio, Fulvio; Recchia, Alessandra
abstract

Preclinical and clinical studies showed that autologous transplantation of epidermis derived from genetically modified epithelial stem cells (EpSCs) leads to long-term correction of inherited skin adhesion defects. These studies were based on potentially genotoxic retroviral vectors. We developed an alternative gene transfer strategy aimed at targeting a “safe harbor” locus, the adeno-associated virus integration site 1 (AAVS1), by zinc-finger nuclease (ZFN)-induced homologous recombination (HR). Delivery of AAVS1-specific ZFNs and a GFP-expressing HR cassette by integration-defective lentiviral (LV) vectors (IDLVs) or adenoviral (Ad) vectors resulted in targeted gene addition with an efficiency of >20% in a human keratinocyte cell line, >10% in immortalized keratinocytes, and <1% in primary keratinocytes. Deep sequencing of the AAVS1 locus showed that ZFN-induced double-strand breaks are mostly repaired by nonhomologous end joining (NHEJ) in primary cells, indicating that poor induction of the HR-dependent DNA repair pathway may be a significant limitation for targeted gene integration. Skin equivalents derived from unselected keratinocyte cultures coinfected with a GFP-IDLV and a ZFN-Ad vector were grafted onto immunodeficient mice. GFP-positive clones were observed in all grafts up to 18 weeks post-transplantation. By histological and molecular analysis, we were able to demonstrate highly efficient targeting of the AAVS1 locus in human repopulating EpSCs.

2012 - A novel human psoriatic skin reconstruct: The role of transit amplifying keratinocytes and of p75 neurotrophin receptor [Abstract in Rivista]
Truzzi, Francesca; Marconi, Alessandra; Lotti, Roberta; Dallaglio, Katiuscia; Palazzo, Elisabetta; Petrachi, Tiziana; Saltari, Annalisa; Pincelli, Carlo
abstract

.

2012 - Apoptotic pathways in the pathogenesis of pemphigus: targets for new therapies [Articolo su rivista]
Lotti, Roberta; Marconi, Alessandra; Pincelli, Carlo
abstract

Pemphigus is a group of rare autoimmune blistering diseases of the skin in which autoantibodies to desmosome cadherins, desmogleins, induce loss of cell-cell adhesion (acantholysis). In addition to steric hindrance and activation of intracellular phosphorylation cascade signaling pathways, apoptosis has been suggested to contribute to the mechanism by which pathogenic IgG induces acantholysis. We review the literature examining the role of apoptosis in pemphigus. Current data recognize a central role of apoptosis in the mechanisms of blister induction. In particular, here we stress the key role of FasL in pemphigus, as it is able to first induce apoptosis, then acantholysis. Being pro-apoptotic molecules important in blister formation, they could represent new specific targets for pemphigus treatment.

2012 - Changes in survivin subcellular localization correlates with different stages of differentiation in normal and cancerous skin [Abstract in Rivista]
K., Dallaglio; Petrachi, Tiziana; Marconi, Alessandra; Morandi, Paolo; Maiorana, Antonino; Pincelli, Carlo
abstract

.

2012 - Definitions and outcome measures for bullous pemphigoid: Recommendations by an international panel of experts [Articolo su rivista]
Murrell, D. F.; Daniel, B. S.; Joly, P.; Borradori, L.; Amagai, M.; Hashimoto, T.; Caux, F.; Marinovic, B.; Sinha, A. A.; Hertl, M.; Bernard, P.; Sirois, D.; Cianchini, G.; Fairley, J. A.; Jonkman, M. F.; Pandya, A. G.; Rubenstein, D.; Zillikens, D.; Payne, A. S.; Woodley, D.; Zambruno, G.; Aoki, V.; Pincelli, C.; Diaz, L.; Hall, R. P.; Meurer, M.; Mascaro, Jr. J. M.; Schmidt, E.; Shimizu, H.; Zone, J.; Swerlick, R.; Mimouni, D.; Culton, D.; Lipozencic, J.; Bince, B.; Grando, S. A.; Bystryn, J. -C.; Werth, V. P.
abstract

Our scientific knowledge of bullous pemphigoid (BP) has dramatically progressed in recent years. However, despite the availability of various therapeutic options for the treatment of inflammatory diseases, only a few multicenter controlled trials have helped to define effective therapies in BP. A major obstacle in sharing multicenter-based evidences for therapeutic efforts is the lack of generally accepted definitions for the clinical evaluation of patients with BP. Common terms and end points of BP are needed so that experts in the field can accurately measure and assess disease extent, activity, severity, and therapeutic response, and thus facilitate and advance clinical trials. These recommendations from the International Pemphigoid Committee represent 2 years of collaborative efforts to attain mutually acceptable common definitions for BP and proposes a disease extent score, the BP Disease Area Index. These items should assist in the development of consistent reporting of outcomes in future BP reports and studies. © 2011 by the American Academy of Dermatology, Inc.

2012 - E-FABP induces differentiation in normal and psoriatic keratinocytes [Abstract in Rivista]
Dallaglio, Katiuscia; Marconi, Alessandra; Truzzi, Francesca; Lotti, Roberta; Palazzo, Elisabetta; Petrachi, Tiziana; Saltari, Annalisa; Pincelli, Carlo
abstract

.

2012 - PC111: a monoclonal anti-Fas Ligand antibody for the treatment of pemphigus [Abstract in Rivista]
Lotti, Roberta; Marconi, Alessandra; D., Katiuscia; Truzzi, Francesca; Petrachi, Tiziana; Pincelli, Carlo
abstract

.

2012 - Role of neurotrophins on dermal fibroblast survival and differentiation [Articolo su rivista]
Palazzo, Elisabetta; Marconi, Alessandra; Truzzi, Francesca; Dallaglio, Katiuscia; Petrachi, Tiziana; P., Humbert; S., Schnebert; E., Perrier; M., Dumas; Pincelli, Carlo
abstract

Neurotrophins (NTs) belong to a family of growth factors that play a critical role in the control of skin homeostasis. NTs act through the low-affinity receptor p75NTR and the high-affinity receptors TrkA, TrkB and TrkC. Here we show that dermal fibroblasts (DF) and myofibroblasts (DM) synthesize and secrete all NTs and express NT receptors. NTs induce differentiation of DF into DM, as shown by the expression of α-SMA protein. The Trk inhibitor K252a, TrkA/Fc, TrkB/Fc or TrkC/Fc chimera prevents DF and DM proliferation. In addition, p75NTR siRNA inhibits DF proliferation, indicating that both NT receptors mediate DF proliferation induced by endogenous NTs. Autocrine NTs also induce DF migration through p75NTR and Trk, as either silencing of p75NTR or Trk/Fc chimeras prevent this effect, in absence of exogenous NTs. Finally, NGF or BDNF statistically increase the tensile strength in a dose dependent manner, as measured in a collagen gel through the GlaSbox device. Taken together, these results indicate that NTs exert a critical role on fibroblast and could be involved in tissue remodelling and wound healing

2012 - Survivin: a dual player in healthy and diseased skin [Articolo su rivista]
Dallaglio, Katiuscia; Marconi, Alessandra; Pincelli, Carlo
abstract

Survivin belongs to the inhibitor of apoptosis (IAP) protein family, and, in addition to the antiapoptotic functions, it also regulates the cell cycle. The survivin gene generates five major isoforms with diverse and opposite functions. Survivin is highly expressed in cancer and in few normal adult tissues, including skin. It is mostly detected in the nucleus of keratinocyte stem cells (KSCs), but it is also expressed in melanocytes and fibroblasts. Survivin isoforms are differentially detected in subpopulations of human keratinocytes, exerting contrasting activities. Survivin has an important role in the regulation of cell cycle in keratinocytes, and it protects these cells from anoikis and UV-induced apoptosis. In melanoma, survivin is abundantly expressed, and its subcellular localization varies depending upon tumor thickness and invasiveness. Survivin overexpression has been shown in squamous cell carcinoma (SCC), and it is also involved in UVB-induced carcinogenesis. The presence of survivin both in the nucleus and in the cytoplasm throughout the epidermal layers of psoriatic lesions suggests the involvement of this protein in the keratinocyte alterations typical of this disease. Additional studies on the expression of survivin isoforms and their subcellular localization in relation to function will confirm the key role of survivin in the skin and will open the field to new therapeutic strategies for many cutaneous conditions

2012 - The p75 neurotrophin receptor triggers the keratinocyte stem-transit amplifying cell transition in normal human epidermis [Abstract in Rivista]
Marconi, Alessandra; Truzzi, Francesca; Lotti, Roberta; Dallaglio, Katiuscia; Palazzo, Elisabetta; Petrachi, Tiziana; Saltari, Annalisa; Pincelli, Carlo
abstract

.

2011 - A novel human psoriatic skin equivalent: the importance of transit amplifying keratinocytes [Abstract in Rivista]
Lotti, Roberta; Truzzi, Francesca; Marconi, Alessandra; Pincelli, Carlo
abstract

Psoriasis is an immune-mediated disease of the skin characterized by keratinocyte hyperproliferation, altered differentiation and resistance to apoptosis. Histologically, psoriasis is represented by augmented thickness of the epidermal compartment of the skin, acanthosis and intense immune cell infiltrate. Psoriasis is thought to be determined by both genetic and environmental components. Psoriasis has long been considered only an immunocyte-mediated disease, but recent data demonstrate the important role of keratinocytes in triggering the disease. Here we sought to develop an in vitro reconstructed skin model that would display the phenotypic and molecular characteristics of psoriatic epidermis in a controlled manner and in the absence of immune cells, providing a tool for the study of keratinocyte biology and allowing the screening of antipsoriatic drugs. Human skin equivalents were generated in the following way: the dermal compartment was a mixture of rat-tail collagen I and human fibroblasts either from healthy adult skin or from patients with psoriasis. Given the importance of transit amplifying (TA) cells in the pathogenesis of psoriasis, we used either TA cells or stem cells isolated from healthy human skin for the epidermal compartment. As a control, we also analysed skin equivalents raised from the total keratinocyte population. The combination of psoriatic fibroblasts and normal TA cells produced the best psoriatic phenotype, with a statistically significant increase of epidermal thickness, areas of acanthosis and expression of psoriatic markers, such as S100A7/psoriasin, K16 and phospho-Stat3 (Tyr705). Moreover, given the absence of p75NTR in psoriatic epidermis and the apoptotic role of p75NTR receptor mainly expressed by TA keratinocytes, we set up skin reconstructs with either p75NTR-positive or p75NTR-depleted TA cells. Skin reconstructs assembled with p75NTR-positive TA keratinocytes were almost negative for psoriatic markers. By contrast, skin reconstructs generated from p75NTR-depleted TA displayed a more psoriasiform phenotype. These results demonstrate the central role of TA cells in psoriasis together with the importance of psoriatic fibroblasts in triggering the psoriatic modifications.

2011 - Improving clinical trial design in psoriasis: Perspectives from the global dermatology community [Articolo su rivista]
P., van de Kerkhof; J., Barker; C. E., Griffiths; A., Menter; C., Leonardi; M., Young; L., Kemeny; Pincelli, Carlo; H. X., Bachelez; A., Katsambas; St X00e5 M., Hle; E. J., Horn; W., Sterry
abstract

Abstract Background: Clinical trials to test investigational drugs for the treatment of chronic plaque psoriasis currently lack standards for comparison of efficacy and safety data.The majority of studies do not address the important need for long-term treatment. Methods: The International Psoriasis Council (IPC) conducted two surveys of its members to assess the need for gold standards, active comparators, and long-term therapy in clinical trials. In Survey 1, 30 participants delivered viewpoints on active comparators for topical therapy (six questions), systemic therapy (nine questions), and continuous versus intermittent therapy (six questions). In Survey 2, 31 participants provided input on gold standards for treatment (five questions), appropriate comparators (four questions), and continuous versus intermittent therapy (six questions). The IPC leadership interpreted the results after each survey. Results: The majority of participants (77% in Survey 1 and 89% in Survey 2) agreed that studies of investigative treatments should include an active comparator. Participants described the most important feature of a gold standard as a treatment that: is widely used and generally accepted (45%); has the best efficacy (42%); and is well tolerated (13%). The majority agreed that gold standards should be dependent on: patient subgroup; location/extent of psoriasis; and psoriasis subtype/morphology. It was also agreed that continuous therapy for more than 3 years is needed for patients with moderate-to-severe plaque psoriasis. We have provided an expert opinion regarding the definition of a gold standard in psoriasis and have also established that no single treatment can be the gold standard across all subgroups and types of the disease. Conclusions: A single gold standard for the treatment of psoriasis does not exist. The complexity and heterogeneity of psoriasis requires different gold standards for the various manifestations of psoriasis and for subgroups of patients reconciling comorbidities. Of note, 17 experts out of 30 selected methotrexate as the most nominated gold standard amongst systemic agents. The experts support the election of an active comparator as one that is most efficacious over just the best in a particular class. In concordance, 87% of respondents agreed that good tolerability is therefore not the lead criterion for selection of an active comparator in favor of effectiveness and broad use. Patients with moderate-to-severe plaque psoriasis require continuous therapy; this statement was supported by 93% of the experts. Reasons for considering long-term therapy included appearance of comorbidities, impairment of quality of life, possibility of relapse, and subjective complaints such as itch, pain, and joint disease.

2011 - Melanoma and the nervous system-Novel pathways mediated by neurotrophins and their receptors [Capitolo/Saggio]
Pincelli, Carlo; Dallaglio, Katiuscia; Truzzi, Francesca
abstract

Le neurotrofine sono una famiglia di proteine sintetizzate e prodotte da molte cellule cutanee, mentre i loro recettori sono espressi nel melanoma. Le neurotrofine stimolano la proliferazione e la migrazione delle cellule di melanoma.

2011 - Neurotrophins in healthy and diseased skin [Articolo su rivista]
Truzzi, Francesca; Marconi, Alessandra; Pincelli, Carlo
abstract

Neurotrophins (NT) belong to a family of structurally andfunctionally related proteins that, depending on the tissuecontext and the receptors involved, promote either neuronalcell survival and differentiation or cell death. NT, and inparticular NGF, were first identified as neurotrophic factorssupporting the synthesis and development of sensory neuronsin the central and peripheral nervous system. it is now widelyaccepted that NT also act as growth factors in non-neuronalcells, including the skin. in the skin, most cell types are ableto secrete and/or to respond to stimulation by NT, creatinga unique network of molecular signaling in the cutaneousmicroenvironment. Moreover, many skin diseases have beenassociated with an involvement of a number of neural factorsincluding NT, but less attention has been given to the role ofNT as growth factors in the development of skin pathologies.This review summarizes currently data on the expression andfunction of NT and their receptors in several cell types in theskin. Moreover it focuses on the role of the skin NT network intwo cutaneous conditions, melanoma and psoriasis where NTare clearly involved.

2011 - Notch protein expression changes in human skin during ageing, keratinocyte differentiation and UVB-exposure [Abstract in Rivista]
Marconi, Alessandra; Palazzo, Elisabetta; Dallaglio, Katiuscia; Truzzi, Francesca; Lotti, Roberta; Petrachi, Tiziana; Pincelli, Carlo
abstract

Epidermal integrity is guaranteed by the presence of keratinocyte stem cells (KSCs) and the correct balance between cell proliferation, differentiation and apoptosis. The microenviroment or “niche” where KSCs reside has a key role in the regulation of these processes and comprise all the factors expressed or released by skin cells. During ageing, epidermal niche changes determine the behavior of KSCs and their progeny (or transit amplifying (TA) cells). Notch proteins (Notch-1, -2, -3, -4) comprise a family of surface receptors which are implicated in maintaining epidermal homeostasis and, for this reason, they are fundamental for the comprehension of the mechanism within epidermal niche in normal condition and during photoageing. First, we have collected skin samples from different age donors: young (Y-under 20 years), adult (A-between 20 and 50 years) and old (O-over 50 years). We have shown that CK15 and CK10 expression don’t change with age, while there is a reduction in Ki67-positive cells and a increasing in the number of epidermal layers expressing involucrin. In culture, keratinocytes present a reduction in proliferation and in CFE in direct ratio to age. In particular, we have shown that TA cells are eventually the most involved. In skin biopsies, Notch-1 expression shown a reduction with age, while Notch-2 presents a different localization. In keratinocyte cell culture, Notch-1 decrease with age, while Notch-2 expression seems to be higher Y cells and constant in A and O cells. Notch-1 and Notch-2 are mainly expressed by TA cells and both proteins present a cytoplasmic localization in KSCs and TA cells, except for Notch-1 which is also present in the nucleus of TA cells. Moreover, both proteins are mainly expressed and activated after stimulus with calcium. Notch-1 activation decrease in confluent and more differentiated keratinocytes from different age, while its expression present a reduction in subconfluent cells and a increase in confluent cells in direct ratio to age. Notch-2 is mainly expressed and activated with confluence in all age class. Furthermore, Notch-1 is strongly activated after stimulus with UVB75 in Y keratinocyte. In addition, it is re-activated in O keratinocytes after stimulus with UVB5 and it is not expressed in more aged keratinocytes with respect to Y ones. Notch-2 is mainly expressed after stimulus with UVB5. This data confirm the role of Notch proteins within epidermal niche and their possible involvement in the mechanisms of photoaging.

2011 - Notch-1 and Notch-2 modulate keratinocyte stem cell viability and differentiation during skin ageing and UVB exposure [Abstract in Rivista]
Palazzo, Elisabetta; Marconi, Alessandra; M., Dumas; S., Schnebert; Dallaglio, Katiuscia; Truzzi, Francesca; Lotti, Roberta; Petrachi, Tiziana; Pincelli, Carlo
abstract

Notch are a family of surface receptors implicated in maintaining epidermal homeostasis. In the epidermis, CK15 and CK10 expression does not change with age, while there is a reduction in Ki67-positive cells and an increased involucrin expression. In culture, keratinocytes display reduced proliferation and a lower colony forming efficiency, as a function of age. Transit amplifying cells appear more affected than stem keratinocytes by ageing. In epidermal sections, Notch-1 expression shows a reduction with age, while Notch-2 is located in the upper layers in Y (under 20-years), in all layers in A (between 20 and 60-years), and predominantly in the basal layer in O (over 60-years). In cultures, Notch-1 activation decreases with age, while Notch-2 seems to be more activated in Y cells than in A and O cells. Notch-1 and Notch-2 are mainly expressed in the cytoplasm of TA cells, Notch-1 being present also in the nucleus. Notch protein inhibition reduces keratinocyte stem cell viability, possibly through survivin downregulation. Inhibiting Notch-1 also induces G1 arrest in keratinocytes at all ages. Notch proteins are activated by calcium, while Notch-1 activation decreases in more differentiated keratinocytes. On the other hand, Notch-2 is activated at cell confluence, in all age groups. Furthermore, Notch-1 is up-regulated upon UVB irradiation (75 mJ/cm2) in Y keratinocytes and is re-activated in O keratinocytes after UVB (5 mJ/cm2). Taken together, these data confirm the role of Notch proteins within epidermal niche and their possible involvement in the mechanisms of photoaging.

2011 - The p75 neurotrophin receptor acts as a "switch on-off" protein in keratinocyte differentiation [Abstract in Rivista]
Palazzo, Elisabetta; Truzzi, Francesca; Marconi, Alessandra; Lotti, Roberta; Dallaglio, Katiuscia; Petrachi, Tiziana; Pincelli, Carlo
abstract

p75 neurotrophin receptor (p75NTR) mediates neurotrophin (NT) signals alone or in combination with the high affinity receptor Trk, also in cell settings outside the nervous system. In the present work, we studied the role of p75NTR during keratinocyte differentiation. p75NTR protein was up-regulated after serum treatment in both confluent and post-confluent keratinocytes, together with the expression of cytokeratin 10 (CK10) and involucrin. Moreover, calcium treatment of subconfluent keratinocytes induced the expression of involucrin, CK10, and the up-regulation of p75NTR, both at the mRNA and at the protein level. Western blotting and confocal analysis showed that p75NTR positive (+) keratinocytes isolated by magnetic cell sorting, expressed less CK10 and CK15, as compared to p75NTR negative (-) cells. p75NTR+ keratinocytes formed a lower number of colonies and proliferated to a lesser extent, as compared to p75NTR- cells. p75NTR is only expressed in the basal keratinocyte layer and is confined to a MIB-1 negative transit amplifying (TA) cell population. p75NTR+ keratinocytes, isolated from TA cells, expressed more survivin and CK15, while displayed less CK10 than p75NTR- TA. Colony forming efficiency was higher in p75NTR+ TA than in p75NTR- TA cells. Moreover, long term proliferation analysis revealed that p75NTR+ TA yielded a higher number of cells than p75NTR- TA. p75NTR retroviral infection of stem cells induced a more differentiated phenotype, with the same features of TA cells. On the other hand, when p75NTR was silenced, calcium treatment failed to induce differentiation in subconfluent keratinocytes, as shown by the absence of involucrin expression. These results suggest that p75NTR+ keratinocytes represent a subpopulation of TA cells, directly derived from stem cells that just started the differentiation process. p75NTR may act as a “switch on-off” protein in stem-TA transition initiating keratinocyte differentiation.

2011 - p75 neurotrophin receptor mediates apoptosis in transit-amplifying cells and its overexpression restores cell death in psoriatic keratinocytes. [Articolo su rivista]
Truzzi, Francesca; Marconi, Alessandra; P., Atzei; M. C., Panza; Lotti, Roberta; Dallaglio, Katiuscia; R., Tiberio; Palazzo, Elisabetta; Vaschieri, Cristina; Pincelli, Carlo
abstract

p75 neurotrophin receptor (p75NTR) belongs to the TNF-receptor superfamily and signals apoptosis in many cell settings. In human epidermis, p75NTR is mostly confined to the transit-amplifying (TA) sub-population of basal keratinocytes. Brain-derived neurotrophic factor (BDNF) or neurotrophin-4 (NT-4), which signals through p75NTR, induces keratinocyte apoptosis, whereas β-amyloid, a ligand for p75NTR, triggers caspase-3 activation to a greater extent in p75NTR transfected cells. Moreover, p75NTR co-immunoprecipitates with NRAGE, induces the phosphorylation of c-Jun N-terminal kinase (JNK) and reduces nuclear factor kappa B (NF-κB) DNA-binding activity. p75NTR also mediates pro-NGF-induced keratinocyte apoptosis through its co-receptor sortilin. Furthermore, BDNF or β-amyloid cause cell death in TA, but not in keratinocyte stem cells (KSCs) or in p75NTR silenced TA cells. p75NTR is absent in lesional psoriatic skin and p75NTR levels are significantly lower in psoriatic than in normal TA keratinocytes. The rate of apoptosis in psoriatic TA cells is significantly lower than in normal TA cells. BDNF or β-amyloid fail to induce apoptosis in psoriatic TA cells, and p75NTR retroviral infection restores BDNF- or β-amyloid-induced apoptosis in psoriatic keratinocytes. These results demonstrate that p75NTR has a pro-apoptotic role in keratinocytes and is involved in the maintenance of epidermal homeostasis.

2010 - A previously unreported function of beta1B integrin isoform in caspase-8-dependent integrin-mediated keratinocyte death [Articolo su rivista]
Lotti, Roberta; Marconi, Alessandra; Truzzi, Francesca; Dallaglio, Katiuscia; Gemelli, Claudia; R. G., Borroni; Palazzo, Elisabetta; Pincelli, Carlo
abstract

Integrins regulate adhesive cell-matrix interactions and mediate survival signals. On the other hand, unligated or free cytoplasmic fragments of integrins induce apoptosis in many cell types (integrin-mediated death). We have previously shown that b1 integrins expression protects keratinocyte stem cells from anoikis, while the role of the b1B integrin isoform has never been clarified. Here we report that suspended keratinocytes undergo apoptosis via the activation of caspase-8, independently of Fas/Fas Ligand system. Indeed, anti-b1 integrin neutralizing antibodies induced apoptosis in short-hairpin-RNA-Fas-Associated-Death-Domain treated cells. Moreover, before and during suspension, caspase-8 directly associated with b1 integrin, that in turn internalized and progressively degraded, shedding the cytoplasmic domain. b1B was expressed only in the cytoplasm in a perinuclear fashion and remained unaltered during suspension. At 24 hrs, as b1A located close to the nucleus, b1B co-localized with b1A and co-immunoprecipitated with caspase-8. Caspase-8 was activated earlier in b1B integrin transfected keratinocytes, and these cells underwent a higher rate of apoptosis than mock cells. By contrast, caspase-8 was not activated in siRNA b1B transfected cells. These results indicate that when b1A is unligated, b1B is responsible for “integrin-mediated death” in human keratinocytes.

2010 - E-FABP mediates human keratinocyte differentiation [Abstract in Rivista]
Dallaglio, Katiuscia; Marconi, Alessandra; Truzzi, Francesca; Lotti, Roberta; Palazzo, Elisabetta; T., Petrachi; T., Bertalot; Pincelli, Carlo
abstract

.

2010 - Keratinocyte Stem Cells: friends and foes [Articolo su rivista]
Pincelli, Carlo; Marconi, Alessandra
abstract

Skin and its appendages provide a protective barrier against the assaults of the environment. To perform its role, epidermis undergoes an ongoing renewal through a balance of proliferation and differentiation/apoptosis called homeostasis. Keratinocyte stem cells reside in a special microenvironment called niche in basal epidermis, adult hair follicle and sebaceous glands. While a definite marker has yet to be detected, data raised part in humans and part in the mouse system, point to a critical role of stem and its progeny transit amplifying cells in epidermal homeostasis. Stem cells are protected from apoptosis and are long-resident in adult epidermis. This renders them more prone to be the origin of skin cancer. In this review, we will outline the main features of adult stem cells in mouse and humans and discuss their fate in relation to differentiation, apoptosis and cancer.

2010 - Neurotrophins stimulate human fibroblast differentiation, migration and contractile strenght [Abstract in Rivista]
Dallaglio, Katiuscia; Palazzo, Elisabetta; Marconi, Alessandra; C., Viennet; M., Dumas; P., Humbert; Pincelli, Carlo
abstract

.

2010 - The p75 neurotrophin receptor is involved in early keratinocyte differentiation [Abstract in Rivista]
Truzzi, Francesca; Marconi, Alessandra; Lotti, Roberta; Dallaglio, Katiuscia; Palazzo, Elisabetta; T., Petrachi; Pincelli, Carlo
abstract

.

2010 - p75NTR mediates apoptosis in transit amplifying (TA) cells and its overexpression resores cell death in psoriatic keratinocytes [Abstract in Rivista]
Truzzi, Francesca; Marconi, Alessandra; Lotti, Roberta; Dallaglio, Katiuscia; Palazzo, Elisabetta; Borroni, Riccardo; Vaschieri, Cristina; R., Tiberio; Pincelli, Carlo
abstract

.

2009 - Anti-Fas Ligand neutralizing antibodies prevent acantholysis both in vitro and in vivo [Abstract in Rivista]
Marconi, Alessandra; Lotti, Roberta; Truzzi, Francesca; Dallaglio, Katiuscia; Borroni, Riccardo; Palazzo, Elisabetta; Y., Kitajima; Pincelli, Carlo
abstract

.

2009 - Apoptolysis: a novel mechanism of skin blistering in pemphigus vulgaris linking the apoptotic pathways to basal cell shrinkage and suprabasal acantholysis [Articolo su rivista]
A. S., Grando; J. C., Bystryn; A., Chernyavsky; M., Frušić Zlotkin; R., Gniadecki; Lotti, Roberta; Y., Milner; M. R., Pittelkow; Pincelli, Carlo
abstract

Understanding the acantholytic pathways leading to blistering in pemphigus vulgaris (PV) is a key to development of novel treatments. A novel paradigm of keratinocyte damage in PV, termed apoptolysis, links the suprabasal acantholytic and cell death pathways to basal cell shrinkage rendering a 'tombstone' appearance to PV lesions. In contrast to apoptolysis, the classic keratinocyte apoptosis mediating toxic epidermal necrolysis causes death and subsequent sloughing of the entire epidermis. Apoptolysis includes five consecutive steps. (1) Binding of autoantibodies to PV antigens. (2) Activation of EGF receptor, Src, mTOR, p38 MAPK and other signalling elements downstream of ligated antigens, elevation of intracellular calcium and launching of the cell death cascades. (3) Basal cell shrinkage due to: (i) collapse and retraction of the tonofilaments cleaved by executioner caspases; and (ii) dissociation of interdesmosomal adhesion complexes caused by phosphorylation of adhesion molecules. (4) Massive cleavage of cellular proteins by activated cell death enzymes leading to cell collapse, and tearing off desmosomes from the cell membrane stimulating secondary autoantibody production. (5) Rounding up and death of acantholytic cells. Thus, the structural damage (acantholysis) and death (apoptosis) of keratinocytes are mediated by the same cell death enzymes. Appreciation of the unifying concept of apoptolysis have several important implications: (i) linking together a number of seemingly unrelated events surrounding acantholysis; (ii) opening new avenues of investigation into the pathomechanism of pemphigus; and (iii) creating new approaches to the treatment of pemphigus based on blocking the signalling pathways and enzymatic processes that lead to blistering.

2009 - Apoptosis precedes desmoglein cleavage and keratinocyte dissociation in pemphigus: anti-Fas Ligand neutralizing antibodies prevent acantholysis both in vitro and in vivo [Abstract in Rivista]
Lotti, Roberta; E., Shu; Truzzi, Francesca; Palazzo, Elisabetta; Marconi, Alessandra; Y., Kitajima; Pincelli, Carlo
abstract

.

2009 - Endogenous survivin modulates survival and proliferation in normal human keratinocytes and during UVB irradiation [Abstract in Rivista]
Marconi, Alessandra; Dallaglio, Katiuscia; Palazzo, Elisabetta; M., Dumas; Truzzi, Francesca; Lotti, Roberta; F., Bonte; Pincelli, Carlo
abstract

.

2009 - Endogenous survivin modulates survival andproliferation in UVB-treated human keratinocytes [Articolo su rivista]
Dallaglio, Katiuscia; Palazzo, Elisabetta; Marconi, Alessandra; M., Dumas; Truzzi, Francesca; Lotti, Roberta; F., Bontè; Pincelli, Carlo
abstract

Survivin is a bi-functional member of inhibitor ofapoptosis protein family, as it is able to both inhibit apoptosisand to regulate cell cycle. We investigated the role of survivin inhuman keratinocytes under normal conditions and during UVBirradiation. Survivin siRNA decreases proliferation and inducesapoptosis in human keratinocytes, in a mode consistent with themitotic catastrophe. Low doses UVB increase survivin expressionat earlier times, while high doses down-regulate survivin level.Low doses UVB induce cell cycle arrest in G2 ⁄ M, while highdoses UVB cause apoptosis. Moreover, overexpression of survivinprotects keratinocytes from UVB-induced apoptosis, and silencingof survivin renders keratinocytes more susceptible to UVBinducedcell death. Finally, survivin siRNA increases UVB-inducedreduction of cell proliferation. Taken together, these resultsindicate that survivin plays a critical role in epidermalhomeostasis in normal conditions and during UVB exposure, withpossible implication in skin carcinogenesis.

2009 - Neurotrophins and their receptors stimulate fibroblast differentiation, migration and tensile strength [Abstract in Rivista]
Palazzo, Elisabetta; Marconi, Alessandra; Truzzi, Francesca; Lotti, Roberta; Dallaglio, Katiuscia; Borroni, Riccardo; M., Dumas; Pincelli, Carlo
abstract

.

2009 - Neurotrophins in healthy and diseased skin [Abstract in Rivista]
Pincelli, Carlo
abstract

.

2009 - Pharmacogenomics in dermatology: from susceptibility genes to personalized therapy [Articolo su rivista]
Pincelli, Carlo; M., Pignatti; Borroni, Riccardo
abstract

A significant proportion of patients with skin disease do not respond to treatment and adverse drug reactions are a common problem. Genetic factors are important determinants for both drug efficacy and toxicity. The fields of pharmacogenetics and pharmacogenomics examine inter-individual variations in the DNA sequence that are related to drug efficacy and toxicity. Here, we present pharmacogenomic data relevant to dermatology and explore the role of dermatologists in identifying patients who may respond to treatment or experience adverse drug reactions

2009 - The skin neurotrophic network in health and disease [Articolo su rivista]
Borroni, Riccardo; Truzzi, Francesca; Pincelli, Carlo
abstract

Neurotrophins (NTs) belong to a family of structurally and functionally related proteins that, depending on the tissue context and the receptors involved, promote neuronal cell survival and differentiation or cell death. NTs also exert important functions in other organs besides the nervous system, including the skin. The presence in the skin of diverse cell types which are able to secrete and/or to respond to stimulation by NTs creates a unique network of molecular signaling in the cutaneous microenvironment. This review summarizes currently available data on the expression and function of NTs and their receptors in several cell types in the skin (namely, keratinocytes, melanocytes and fibroblasts). The role of the skin NT network in the development and maintenance of some relevant skin diseases is presented and the potential implications for therapeutic intervention are discussed.

2009 - What's New in Laboratory Research? [Articolo su rivista]
Pincelli, Carlo; Lotti, Roberta
abstract

Psoriasis is a multigenic disease with a number of susceptibility loci on different chromosomes predisposing to the disease, which is in turn triggered by environmental factors. The pathogenesis of psoriasis is characterized by 2 main components, the dysfunctions of the immune system and the alteration of keratinocyte homeostasis. While the Th1 T cell response has long been considered the sole immune agent in the pathomechanisms of psoriasis, recently, the role of IL-23-driven Th17 has been shown to be predominant. Subsequently, only effector memory T cells expressing alpha1beta1 integrin migrate into the epidermis, and psoriasis development is inhibited by blocking this integrin. Psoriatic epidermis contains high amounts of antimicrobial peptides, which have been shown to be a key mediator of plasmocytoid dendritic cell activation in psoriasis. The keratinocyte component has recently regained the central stage, as the abrogation of JunB/activator protein 1 in mouse keratinocytes induces development of psoriasiform lesions in T cell deficient mice. Moreover, inhibition of nerve growth factor and its receptor in keratinocytes strikingly improves psoriatic lesions.

2008 - Defective p75 neurotrophin receptor in transit amplifying cells renders psoriatic keratinocytes more resistant to apoptosis [Abstract in Rivista]
Truzzi, Francesca; Marconi, Alessandra; Lotti, Roberta; Dallaglio, Katiuscia; Giannetti, Alberto; Pincelli, Carlo
abstract

.

2008 - Inhibition of keratinocyte proliferation with a novel compound for the topical treatment of psoriasis and dermatitis [Abstract in Rivista]
L., Bertarione Rava Rossa; S., Traversa; V., Mainero; Pincelli, Carlo; Marconi, Alessandra
abstract

.

2008 - Neurothrophins and their receptors stimulate melanoma cell proliferation and migration [Abstract in Rivista]
Truzzi, Francesca; Marconi, Alessandra; Lotti, Roberta; Dallaglio, Katiuscia; Borroni, Riccardo; L., French; Pincelli, Carlo
abstract

.

2008 - Neurotrophins and their receptors stimulate melanoma cell proliferation and migration [Articolo su rivista]
Truzzi, Francesca; Marconi, Alessandra; Lotti, Roberta; Dallaglio, Katiuscia; L. E., French; B. L., Hempstead; Pincelli, Carlo
abstract

Melanoma is a highly aggressive skin tumor that originates in the epidermis from melanocytes. As melanocytes share with the nervous system a common neuroectodermal origin and express all neurotrophins (NTs), we evaluated the expression and function of NTs and their receptors in melanoma. We report that primary and metastatic melanoma cell lines synthesize and secrete all NTs. Moreover, melanoma cells express the low-affinity (p75NTR) and the high-affinity tyrosine kinase NT receptors (Trk). The inhibition of Trk receptors by either K252a or Trk/Fc chimeras prevents proliferation, indicating that autocrine NTs are responsible for this effect. NT-3, NT-4, and nerve growth factor (NGF) induce cell migration, with a stronger effect on metastatic cell lines. Transfection with p75NTR small interfering RNA (p75NTRsiRNA) or treatment with K252a inhibits NT-induced melanoma cell migration, indicating that both the low- and high-affinity NT receptors mediate this effect. All melanoma cell lines express the p75NTR coreceptor sortilin by which proNGF stimulates migration in melanoma cells, but not in cells transfected with p75NTRsiRNA. These results indicate that NTs, through their receptors, play a critical role in the progression of melanoma.

2008 - P75 neurotrophin receptor restores defective apoptosis in psoriatic keratinocytes: a role for transit amplifying cells [Abstract in Rivista]
Marconi, Alessandra; Truzzi, Francesca; Lotti, Roberta; Dallaglio, Katiuscia; Borroni, Riccardo; Vaschieri, Cristina; R., Tiberio; Pincelli, Carlo
abstract

.

2008 - Pathologic changes after photodynamic therapy for basal cell carcinoma and Bowen disease: A histologic and immunohistochemical investigation [Articolo su rivista]
F., Fantini; A., Greco; A. M., Cesinaro; T., Surrenti; K., Peris; Vaschieri, Cristina; Marconi, Alessandra; Giannetti, Alberto; Pincelli, Carlo
abstract

Objective: To investigate the in vivo reactions and themechanisms of cell death after photodynamic therapy(PDT) for cutaneous carcinomas. Photodynamic therapyis a new treatment modality for nonmelanoma skin cancers.Its effects on target tissue have been well investigatedin vitro, where apoptosis appears to be the maineffector mechanism, but its effects remain undefined invivo.Design: Skin biopsy specimens were obtained sequentiallyafter PDT for basal cell carcinoma and in situ squamouscell carcinoma (Bowen disease). Evidence from routinehistologic evaluation was compared with a panel ofapoptosis-related (TUNEL [terminal deoxynucleotidyltransferase-mediated biotin–deoxyuridine triphosphatenick-end labeling], caspase-3, and Bcl-2) and inflammatory(CD4, CD8, CD20, CD68, and CD56) markers. Weused electron microscopy to evaluate cell damage at theultrastructural level.Main Outcome Measures: Evidence of the mechanismsof tumor cell damage after PDT, detection of histologicand/or immunohistochemical signs of apoptosis,and time course of the tumor destruction andinflammatory reaction.Results: Early epidermal damage and an acute dermalinflammatory response were detected 15 minutes afterPDT. In basal cell carcinoma, nodule damage progressedfrom scant apoptotic cells seen at the dermalepithelialjunction to massive destruction seen after 1 and2 days. The periphery of the basaloid nodules consistentlyshowed earlier and predominant damage, as demonstratedby the perfect coincidence of histologic and immunohistochemicalevidence with apoptotic markers(TUNEL and caspase-3 staining). Fibrosis and lentigolikechanges were seen in late biopsy specimens.Conclusions: This study defines the time course and characteristicsof the skin tumor response to PDT. Taken together,our observations suggest that direct damage tocancer cells is the main effector mechanism leading toPDT response. The involvement of apoptosis is demonstratedby the simultaneous appearance of histologic, immunohistochemical,and ultrastructural markers that occurin the early phases of the cutaneous reaction to PDT.These observations could help to develop future refinementsof the PDT technique.

2008 - Pin Cell [Spin Off]
Pincelli, Carlo; Marconi, Alessandra
abstract

PinCell is a biotech company founded in October 2008 as an academic spin-off of the University of Modena and Reggio Emilia by initiative of Prof. Carlo Pincelli and Dr. Alessandra Marconi. The Mission is to research and develop novel molecules that will increase the therapeutic options for neoplastic and chronic inflammatory skin diseases. The research and development process is focused on drug discovery, drug physico-chemical characterization, preclinical pharmacology and toxicology studies and early phase clinical trials on healthy volunteers (phase I) PinCell directs its discoveries to other biotechnological companies or to pharmaceutical industries in need for novel ideas for drug development or looking for products at advanced stage of development requiring rapid commercial transfer.

2008 - Remedies for pemphigus containing anti Fas ligand antibodies [Brevetto]
Pincelli, Carlo; Marconi, Alessandra
abstract

The present invention refers to the use of FasL antagonists, e.g. of humanized antibodies directed against human Fas ligands (also named CD95L or Apo1L and hereinafter abbreviated as FasL) for the prevention and/or treatment of skin diseases associated with keratinocytes acantholysis, particularly for the prevention and/or treatment of pemphigus.

2008 - Survivin siRNA causes cell cycle arrest in G2/M and increases UVB-induced apoptosis in human keratinocytes [Abstract in Rivista]
Dallaglio, Katiuscia; Marconi, Alessandra; Lotti, Roberta; Truzzi, Francesca; M., Dumas; F., Bonte; Pincelli, Carlo
abstract

.

2007 - A novel topical treatment of psoriasis: inhibition of NGF-induced keratinocyte proliferation [Abstract in Rivista]
L., Bertarione Rava Rossa; S., Traversa; V., Mainero; D., Barone; C., Oderda; S., Fumero; Pincelli, Carlo; Marconi, Alessandra
abstract

.

2007 - CT327: a novel mini-PEGylated compound for topical treatment of psoriasis [Abstract in Rivista]
L., Bertarione Rava Rossa; S., Traversa; V., Mainero; D., Barone; C., Oderda; S., Fumero; Pincelli, Carlo; Marconi, Alessandra
abstract

.

2007 - CT327: a novel topical treatment of psoriasis and dermatitis [Abstract in Rivista]
L., Bertarione Rava Rossa; S., Traversa; V., Mainero; D., Barone; C., Oderda; S., Fumero; Pincelli, Carlo; Marconi, Alessandra
abstract

.

2007 - Carboxyfullerenes localize within mitochondria and prevent the UVB-induced intrinsic apoptotic pathway [Articolo su rivista]
F., Chirico; C., Fumelli; Marconi, Alessandra; A., Tinari; E., Straface; W., Malorni; R., Pellicciari; Pincelli, Carlo
abstract

Carboxyfullerenes (CF) act as free radical scavengers in many cell settings and prevent apoptosis in vitro and in vivo. CF protect normal human keratinocytes from UVB-induced apoptosis, although the mechanisms underlying this effect remain to be clarified. Double-staining confocal laser microscopy revealed that CF penetrate the cell and colocalize with cytokeratin-18 within cytoplasm. This localization was confirmed by transmission electron microscopy that showed CF intermingled with keratin filaments. Moreover, double-staining with the mitochondrial marker anti-F1-ATPase antibody demonstrated that CF are expressed in mitochondria. Transmission electron microscopy confirmed that CF actually localize within mitochondria. Then, normal human keratinocytes were UVB-irradiated in the presence or absence of CF at different doses. CF protected keratinocytes from apoptosis induced by reactive oxygen species. CF scavenging effect is associated with a partial blockade of the UVB-induced intrinsic apoptotic pathway by down-modulating caspase-9 activation and cytochrome c release, and by inhibiting the down-regulation of the inhibitor of apoptosis proteins (IAP) survivin, livin, IAP-1 and IAP-2. Finally, CF prevented the cleavage of Bid, up-regulation of Bad and down-regulation of Mcl-1 induced by UVB. Taken together, these results indicate that CF penetrate human keratinocytes, localize within mitochondria where they act both by scavenging free radicals and by protecting cells from apoptosis.

2007 - Fas ligand and pemphigus sera induce desmoglein cleavage and apoptosis in human keratinocytes [Abstract in Rivista]
Lotti, Roberta; Marconi, Alessandra; Truzzi, Francesca; Dallaglio, Katiuscia; Vaschieri, Cristina; Pincelli, Carlo
abstract

Pemphigus is an autoimmune bullous disease characterized by loss of adhesion of keratinocytes that round up in a process known as acantholysis. While the molecular mechanisms underlying acantholysis are still unclear, it is well known that cell detachment is often associated with apoptosis. We have previously detected apoptotic keratinocytes in perilesional, yet undetached, pemphigus skin. Moreover, we have observed that Fas ligand (FasL) levels are significantly higher in untreated pemphigus sera (more than 0.1 ng/ml) than in controls. As pemphigus sera induce apoptosis in cultured human keratinocytes, we wanted to analyze the apoptotic mechanisms in pemphigus. We first confirmed that pemphigus lesions contain apoptotic keratinocytes, by showing caspase-3 activation. In addition, while Fas receptor (FasR) is expressed in the basal and partially in the suprabasal layers in pemphigus vulgaris (PV), it is detected throughout the epidermal layers in muco-cutaneous pemphigus. Furthermore, pemphigus sera-induced keratinocyte apoptosis is partially prevented by pretreatment with either caspase-8 inhibitor or anti-FasL neutralizing antibody. Moreover, caspase-8 activation induced by untreated pemphigus sera is partially inhibited by anti-FasL antibody. Untreated pemphigus sera induce the cleavage of desmoglein 1 and 3 (dsg 1, 3). Moreover, recombinant FasL dose-dependently cleaves dsg 1 and 3 (0.1, 10, 100 ng/ml). Finally, caspase-8 inihibitor prevents dsg cleavage, strongly indicating the critical role of FasL in the pathogenesis of pemphigus. In particular, high levels of FasL, contained in pemphigus sera exert a dual activity, by both inducing keratinocyte apoptosis and dsg cleavage.

2007 - Neurotrophins act as neuroendocrine regulators of skin homeostasis in health and disease [Articolo su rivista]
E. M. J., Peters; U., Raap; P., Welker; A., Tanaka; H., Matsuda; S., Pavlovic Masnicosa; S., Hendrix; Pincelli, Carlo
abstract

Neurotrophins regulate cutaneous innervation, act as growth and motility factors on structural skin cells such as keratinocytes and fibroblasts, modulate cutaneous immune function and even serve as stress mediators in skin biology. The multilayered neurotrophin interaction with skin biology through high affinity specific tyrosinekinase receptors and the Janus-faced p75 receptor, which depending on ligand and co-receptor expression can serve as a low-affinity pan-neurotrophin receptor or a high affinity proneurotrophin receptor, guaranties this neuroendocrine peptide family a central position in the control of skin homeostasis in health and disease. It is a challenging task for future research efforts to integrate our knowledge on differential neurotrophin expression patterns and signaling pathways into complex concepts of neuroendocrine tissue remodeling and pathogenetic processes. In addition, we need to improve our understanding of the role of neurotrophin processing enzymes, associated co-receptors and intracellular adaptor molecules in specific cutaneous cell populations to design precise interaction tools for research and treatment. Such tools will allow us to utilize this ancient growth factor family in the management of neurotrophin responsive pathogenetic pathways and cutaneous diseases such as neurogenic inflammation, peripheral nerve degeneration, wound healing, atopic dermatitis or psoriasis.

2007 - P75 neurotrophin receptor (NTR) mediates chemotherapy-induced apoptosis in melanoma cells [Abstract in Rivista]
Truzzi, Francesca; Marconi, Alessandra; Lotti, Roberta; Dallaglio, Katiuscia; Borroni, Riccardo; Pincelli, Carlo
abstract

.

2007 - P75 neurotrophin receptor exerts pro-apoptotic functions in human keratinocytes: a role in psoriasis [Abstract in Rivista]
Marconi, Alessandra; Truzzi, Francesca; Lotti, Roberta; Dallaglio, Katiuscia; Vaschieri, Cristina; R., Tiberio; Pincelli, Carlo
abstract

.

2007 - Remedies for pemphigus containing anti Fas Ligand antibodies [Brevetto]
Pincelli, Carlo; Marconi, Alessandra
abstract

Disclosure of the use of FasL antagonist, e.g. of humanized antibodies directed against human Fas Ligands (also named CD95L or Apo1L and hereinafter abbreviated as FasL) for the prevention and/or treatment of skin diseases associated with keratinocytes acantholysis, particularly for the prevention and/or treatment of pemphigus

2007 - Survivin identifies keratinocyte stem cells and is downregulated by anti-β1 integrin during anoikis [Articolo su rivista]
Marconi, Alessandra; Dallaglio, Katiuscia; Lotti, Roberta; Vaschieri, Cristina; Truzzi, Francesca; F., Fantini; Pincelli, Carlo
abstract

Survivin belongs to the family of inhibitor of apoptosis proteins and is involved in regulation of cell death as well as cell division. Here, we show that wild-type (WT) survivin is expressed in a subpopulation of basal keratinocytes in normal human skin at the cytoplasmic level. WT survivin is highly expressed in keratinocyte stem cells (KSCs), whereas its mRNA level decreases in transit amplifying (TA) cells and disappears in postmitotic (PM) cells. Likewise, WT survivin protein is expressed in KSCs, almost undetectable in TA cells, and absent in PM cells. Real time polymerase chain reaction demonstrates that the putative antiapoptotic isoforms survivin-2B and survivin-Delta Ex3 are expressed at the highest levels in KSCs, whereas they tend to decrease in TA cells and disappear in PM cells. On the contrary, the putative proapoptotic variants of survivin, survivin-3B, and survivin-2 alpha tend to be high in PM and TA cells and are almost absent in KSCs. By confocal microscopy, survivin is predominantly expressed at the nuclear level in KSCs, which proliferate significantly better than TA cells, which, in turn, express mostly cytosolic WT survivin. Blocking beta 1 integrin signal downregulates WT survivin mRNA and protein expression and induces apoptosis (anoikis) in KSCs. On the other hand, inhibition of beta 1 integrin upregulates mRNA expression of survivin-2 alpha. Taken together, these results indicate that survivin identifies human KSCs. Expression of nuclear survivin could reflect the different behavior between KSCs in vitro and in vivo, in terms of proliferation. Finally, survivin could be part of the niche protection by preventing anoikis in KSCs.

2007 - Survivin modulates cell cycle and interferes with UV-induced apoptosis in human keratinocytes [Abstract in Rivista]
Dallaglio, Katiuscia; Marconi, Alessandra; M., Dumas; F., Bonte; Lotti, Roberta; Truzzi, Francesca; Gemelli, Claudia; Pincelli, Carlo
abstract

.

2007 - The incidence of arthropathy adverse events in efalizumab-treated patients is low and similar to placebo and does not increase with long-term treatment: pooled analysis of data from Phase III clinical trials of efalizumab. [Articolo su rivista]
Pincelli, Carlo; E., Henninger; F., Casset Semanaz
abstract

A large-scale, pooled analysis of safety data from five Phase III clinical trials (including open-label extensions of two of these studies) and two Phase III open-label clinical trials of efalizumab was conducted to explore whether arthropathy adverse events (AEs) were associated with efalizumab treatment in patients with moderate-to-severe chronic plaque psoriasis. Data from patients who received subcutaneous injections of efalizumab or placebo were stratified for analysis into phases according to the nature and duration of treatment. These included: the 'first treatment' phase (0-12- week data from patients who received either efalizumab, 1 mg/kg once weekly, or placebo in the five placebo-controlled studies); the 'extended treatment' phase (13-24-week data from seven trials for all efalizumab-treated patients); and the 'long-term treatment' phase (data from efalizumab-treated patients who received treatment for up to 36 months in two long-term trials). Descriptive statistics were performed and the incidence of arthropathy AEs per patient-year was calculated using 95% confidence intervals (CIs). During the first treatment phase, a similar proportion of patients had an arthropathy AE in the efalizumab group (3.3%; 58/1740 patients) compared with the placebo group (3.5%; 34/979 patients); the incidence of arthropathy AEs per patient-year was 0.15 in the efalizumab group (95% CI 0.11-0.19) and 0.16 in the placebo group (95% CI 0.11-0.22). Analysis of first treatment phase data from one study (n = 793) showed that the incidence of psoriatic arthropathy per patient-year was lower in efalizumab-treated patients (0.10; 95% CI 0.05-0.18) than in those given placebo (0.17; 95% CI 0.08-0.30). During the extended treatment phase, the incidence of arthropathy remained low (0.17; 95% CI 0.14-0.22). Data from two long-term studies showed that there was no increase in the incidence of arthropathy AEs over time in patients treated with efalizumab for up to 36 months. Patients who had an arthropathy AE during treatment with efalizumab appeared to be more likely to have a history of arthropathy prior to treatment. Efalizumab does not appear to increase the risk of arthropathy AEs compared with placebo.

2007 - p75 neurotrophin receptor (NTR) is up-regulated by chemotherapy and induces apoptosis in melanoma cells [Abstract in Rivista]
Marconi, Alessandra; Truzzi, Francesca; Lotti, Roberta; Dallaglio, Katiuscia; Pincelli, Carlo
abstract

.

2006 - Anoikis is triggered by the association between beta 1B integrin and caspase-8 in human keratinocytes [Abstract in Rivista]
Lotti, Roberta; Marconi, Alessandra; Dallaglio, Katiuscia; Truzzi, Francesca; Pincelli, Carlo
abstract

.

2006 - Apoptosis and skin tumors [Abstract in Rivista]
Pincelli, Carlo
abstract

.

2006 - Apoptosis in pemphigus [Abstract in Rivista]
Pincelli, Carlo
abstract

.

2006 - Are desmoglein autoantibodies essential for the immunopathogenesis of pemphigus vulgaris, or just "witnesses of disease"? [Articolo su rivista]
M., Amagai; A. R., Ahmed; Y., Kitajima; J. C., Bystryn; Y., Milner; R., Gniadecki; M., Hertl; Pincelli, Carlo; H., Kurzen; M., Fridkis Hareli; Y., Aoyama; M., Frušić Zlotkin; E., Müller; M., David; D., Mimouni; D., Vind Kezunovic; B., Michel; M., Mahoney; S., Grando
abstract

.

2006 - Biologicals in the treatment of psoriasis [Articolo su rivista]
Pincelli, Carlo; Giannetti, Alberto
abstract

.

2006 - Efalizumab therapy and incidence of arthropathy adverse events: analysis of pooled data from phase II/III/IV clinical trials [Abstract in Rivista]
Pincelli, Carlo; F., Casset Semanaz
abstract

.

2006 - Expression and function of neurotrophins and their receptors in human melanocytes. [Articolo su rivista]
Marconi, Alessandra; Panza, Mc; Bonnet Duquennoy, M; Lazou, K; Kurfurst, R; Truzzi, Francesca; Lotti, Roberta; DE SANTIS, Giorgio; Dumas, M; Bonté, F; Pincelli, Carlo
abstract

Melanocytes and cells of the nervous system are of common ectodermal origin and neurotrophins (NT) have been shown to be released by human keratinocytes. We investigated the expression and function of NT [nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), NT-3, NT-4/-5] and their receptors in human melanocytes. Human melanocytes produce all NT in different amounts, whereas they only release NT-4. NT-4 release is downregulated, whereas NT-3 is upregulated by ultraviolet (UVB) irradiation. Melanocytes treated with phorbol 12-myristate 13-acetate (PMA) express TrkA and TrkB, but not TrkC. NT fail to stimulate melanocyte proliferation, whereas they stimulate the synthesis of tyrosinase and tyrosinase-related protein-1 (TRP-1). Finally, NT-3, NT-4 and NGF increase melanin production. Taken together, these results demonstrate an intriguing interaction between melanocytes and the nervous system. We speculate that NT could be considered the target of therapy for disorders of skin pigmentation.

2006 - Fas ligand and pemphigus sera induce cleavage of desmogleins and apoptosis in human keratinocytes [Abstract in Rivista]
Marconi, Alessandra; Truzzi, Francesca; Lotti, Roberta; Vaschieri, Cristina; Dallaglio, Katiuscia; Giannetti, Alberto; Pincelli, Carlo
abstract

.

2006 - Imiquimod modulates the expression of survivin, bcl-2 and Ki67 in skin tumors [Abstract in Rivista]
Marconi, Alessandra; Dallaglio, Katiuscia; Lotti, Roberta; Vaschieri, Cristina; K., Peris; Pincelli, Carlo
abstract

.

2006 - Neuropeptides and Atopic Eczema [Capitolo/Saggio]
F., Fantini; Pincelli, Carlo
abstract

.

2006 - Neurotrophin modulate melanogenesis in human melanocytes [Abstract in Rivista]
C., Lazou; Marconi, Alessandra; C., Marteau; M., Bonnet Duquennoy; R., Kurfurst; M., Dumas; F., Bonte; Pincelli, Carlo
abstract

.

2006 - Neurotrophins (NT) modulate survival and chemotaxis in metastatic and primary melanoma cell lines through their high-affinity receptors Trk [Abstract in Rivista]
Truzzi, Francesca; Marconi, Alessandra; Lotti, Roberta; Dallaglio, Katiuscia; M., Pignatti; L., French; Pincelli, Carlo
abstract

.

2006 - Neurotrophins (NT) modulate survival and chemotaxis in metastatic and primary melanoma cell lines through their high-affinity receptors Trk [Abstract in Rivista]
Truzzi, Francesca; Marconi, Alessandra; Lotti, Roberta; Dallaglio, Katiuscia; M., Pignatti; L., French; Pincelli, Carlo
abstract

.

2006 - Neurotrophins in skin biology and pathology [Articolo su rivista]
V. A., Botchkarev; M., Yaar; E. M. J., Peters; S. P., Raychaudhuri; N. V., Botchkareva; Marconi, Alessandra; S. K., Raychaudhuri; R., Paus; Pincelli, Carlo
abstract

Neurotrophins (NTs) belong to a family of growth factors, which control the development, maintenance, and apoptotic death of neurons and also fulfill multiple regulatory functions outside the nervous system. Biological effects induced by NTs strongly depend on the pattern of NT receptor/ co-receptors expression in target cells, as well as on the set of intracellular adaptor molecules that link NT signalling to distinct biochemical pathways. In this review, we summarize data on the molecular mechanisms underlying the involvement of NTs in the control of non- neuronal functions in normal skin ( e. g. keratinocyte proliferation, melanocyte development and apoptosis, hair growth). We also review the data on the role for NTs and their receptors in a number of pathological skin conditions ( stress- induced hair loss, psoriasis, atopic dermatitis). Although additional efforts are required to fully understand mechanisms underlying the involvement of NTs and their receptors in controlling functions of normal and pathologically altered skin cells, substantial evidence suggests that modulation of NT signalling by NTs receptor agonists/ antagonists may be developed as intervention modalities in distinct skin and hair growth pathologies.

2006 - Strategy for in vitro and ex-vivo testing of neutrophins signaling pathways based on immunoproteomic array profiling [Abstract in Rivista]
Atzei, Paola; X., Huang; M. C., Fargnoli; M., Krajewska; Pincelli, Carlo; K., Peris; J. C., Reed; S. I., Matsuzawa; S., Krajewski
abstract

.

2006 - Survivin identifies keratinocyte stem cells and it is down-regulated by anti beta 1 integrin during anoikis [Abstract in Rivista]
Dallaglio, Katiuscia; Lotti, Roberta; Marconi, Alessandra; Truzzi, Francesca; M., Dumas; F., Bonte; Pincelli, Carlo
abstract

.

2006 - p75 Neurotrophin Receptor (p75NTR) signals apoptosis either alone or in association with its co-receptor sortilin in human keratinocytes [Abstract in Rivista]
Truzzi, Francesca; Marconi, Alessandra; Atzei, Paola; Lotti, Roberta; Dallaglio, Katiuscia; Pincelli, Carlo
abstract

.

2005 - Apoptotic mechanisms of acantholysis in pemphigus [Abstract in Rivista]
Pincelli, Carlo
abstract

.

2005 - Beta 1 integrin associates with caspase-8 and triggers anoikis in human keratinocytes [Abstract in Rivista]
Lotti, Roberta; Marconi, Alessandra; Panza, Maria Cristina; Pincelli, Carlo
abstract

.

2005 - Blockade of beta 1 integrin directly activates caspase-8-triggered extrinsic apoptotic pathway in human keratinocytes [Abstract in Rivista]
Lotti, Roberta; Marconi, Alessandra; Panza, Maria Cristina; Gemelli, Claudia; M., Leverkus; Giannetti, Alberto; Pincelli, Carlo
abstract

.

2005 - Brain-derived neurotrophic factor (BDNF) and pro-nerve growth factor (NGF) induce apoptosis in human keratinocytes through the p75 neurotrophin receptor [Abstract in Rivista]
Panza, Maria Cristina; Atzei, Paola; Truzzi, Francesca; Lotti, Roberta; Marconi, Alessandra; Pincelli, Carlo
abstract

.

2005 - Evaluation of different serological tests for the diagnosis of pemphigus [Abstract in Rivista]
Vaschieri, Cristina; Marconi, Alessandra; Pincelli, Carlo; Giannetti, Alberto
abstract

.

2005 - POLYMER CONJUGATES OF K-252A AND DERIVATIVES THEREOF [Brevetto]
Traversa, S.; Bagnod, R.; Barone, D.; Beratarione Rava Rossa, L.; Fumero, S.; Mainero, V.; Marconi, A.; Oderda, C.; Pincelli, C.; Lorenzetto, C.; Beccaria, L.
abstract

The present invention relates to novel polymer conjugates of K-252a and derivatives thereof and to their use for the preparation of a pharmaceutical composition useful for the prevention, alleviation and treatment of kinase-associated pathologies. In particular, the present invention relates to the prevention, alleviation and treatment of HMGB1-associated pathologies. In a particular aspect, the invention relates to the use of the novel polymer conjugates of K-252a and derivatives thereof in the preparation of a pharmaceutical composition useful for the prevention, alleviation and treatment of neurological disorders, neuropathies and neurodegenerative disorders of the central and peripheral nervous system.; In a further preferred aspect, the invention relates to the use of the polymer conjugates in the preparation of a pharmaceutical composition useful for the prevention, alleviation and treatment of dermal pathologies, in particular dermal pathologies associated with an excessive keratinocyte proliferation, in particular psoriasis. In a still further aspect, the invention relates to the use of the polymer conjugates in the prevention, alleviation and treatment of NGF-related pain. More specifically, the present invention relates to a polymer conjugate of K-252a and derivatives thereof, wherein the polymer is polyethylene glycol or methoxy-polyethylene glycol formula.

2005 - Phenotype and function of a keratinocyte subpopulation enriched in stem cells. [Abstract in Rivista]
Marconi, Alessandra; Panza, Maria Cristina; Lotti, Roberta; M., Dumas; F., Bontè; Pincelli, Carlo
abstract

.

2005 - Survivin is overexpressed in keratinocyte stem cells and is down-regulated by pro-apoptotic agents [Abstract in Rivista]
Marconi, Alessandra; Panza, Maria Cristina; Lotti, Roberta; Vaschieri, Cristina; K., Peris; Pincelli, Carlo
abstract

.

2005 - p75 neurotrophin receptor co-localizes with sortilin and induces apoptosis in a subpopulation of human keratinocytes [Abstract in Rivista]
Atzei, Paola; Marconi, Alessandra; Panza, Maria Cristina; Vaschieri, Cristina; Truzzi, Francesca; J. C., Reed; Pincelli, Carlo
abstract

.

2004 - Blockade of beta(1) integrin activates caspase-8 apoptotic pathway without Fas/FasL modulation in human keratinocytes [Abstract in Rivista]
Marconi, Alessandra; Lotti, Roberta; Panza, Maria Cristina; T., Wachter; M., Leverkus; Giannetti, Alberto; Pincelli, Carlo
abstract

.

2004 - Carboxyfullerenes localize at the intracellular level and prevents caspase cascade induced by UVB in human keratinocytes [Abstract in Rivista]
C., Fumelli; Marconi, Alessandra; M., Pignatti; Vaschieri, Cristina; Giannetti, Alberto; Pincelli, Carlo
abstract

.

2004 - Effect of NGF on matrix metalloproteinases and tissue inhibitors of metalloproteinases produced by human dermal fibroblasts and myofibroblasts [Abstract in Rivista]
C., Gondran; M., Dumas; Marconi, Alessandra; Truzzi, Francesca; Atzei, Paola; F., Bonte; Pincelli, Carlo; S., Schnebert
abstract

.

2004 - FLICE/caspase-8 activation triggers anoikis induced by beta(1)-integrin blockade in human keratinocytes [Abstract in Rivista]
Marconi, Alessandra; Fila, Chiara; Panza, Maria Cristina; R., Tiberio; T., Wacter; M., Leverkus; Pincelli, Carlo
abstract

.

2004 - FLICE/caspase-8 activation triggers anoikis induced by beta(1)-integrin blockade in human keratinocytes [Articolo su rivista]
Marconi, Alessandra; P., Atzei; C., Panza; C., Fila; R., Tiberio; Truzzi, Francesca; T., Wachter; M., Leverkus; Pincelli, Carlo
abstract

Beta(1)-integrin protects keratinocyte stem cells (KSC) from cell-detachment apoptosis ('anoikis'). Here we show that caspase-8 active protein is detected in both young transit amplifying (TA) cells and TA cells, but not in KSC. On suspension, caspases are activated earlier in young TA than in KSC, whereas anti-beta(1)-integrin neutralizing antibody accelerates caspase activation in both KSC and young TA. Caspases 8 and 10 are the first caspases to be activated whereas caspase-8 inhibitor zIETD-fmk delays the activation of Bid, caspase-9 and caspase-3. However, the caspase-9 inhibitor zLEDH-fmk does not block the activation of caspase-8, Bid, caspase-10 and caspase-3. Moreover, caspase-8, but not caspase-9 inhibitor partially prevents keratinocyte anoikis. As FLIP inhibits caspase-8 processing, we retrovirally infected HaCaT keratinocytes with c-FLIPL. Anti-beta(1)-integrin fails to activate caspase-8, Bid, caspase-9 and to induce the release of cytochrome c in c-FLIPL overexpressing keratinocytes. Finally, overexpression of c-FLIPL partially prevents anoikis in both suspended and anti-beta(1) integrin-treated cells. Taken together, these results indicate that the extrinsic apoptotic pathway triggered by caspase-8 predominates in keratinocyte anoikis. However, the release of cytochrome c and the later activation of caspase-9 seem to suggest that the intrinsic mitochondrial pathway may intervene as a positive feedback loop of caspase activation.

2004 - Nerve growth factor and its role in epidermal homeostasis [Abstract in Rivista]
Pincelli, Carlo
abstract

.

2004 - Nerve growth factor receptors modulates apoptosis in melanoma cell lines [Abstract in Rivista]
Truzzi, Francesca; Marconi, Alessandra; Atzei, Paola; M., Pignatti; L., French; Pincelli, Carlo
abstract

.

2004 - The neurotrophin network in human skin [Abstract in Rivista]
Marconi, Alessandra; M., Dumas; Fila, Chiara; Truzzi, Francesca; Atzei, Paola; M., Pignatti; F., Bonte; Giannetti, Alberto; Pincelli, Carlo
abstract

.

2004 - The p75 neurotrophin receptor signals apoptosis in a subpopulation of human keratinocytes [Abstract in Rivista]
Atzei, Paola; Marconi, Alessandra; M., Pignatti; Truzzi, Francesca; Giannetti, Alberto; Pincelli, Carlo
abstract

.

2003 - Apoptosis in prostate carcinoma before and after neoadjuvant therapy with LH-RH analog [Articolo su rivista]
S., Bencini; Migaldi, Mario; G., Castagnetti; Pincelli, Carlo; P., Ferrari; DE GAETANI, Carmela; G. P., Trentini
abstract

This study investigated apoptosis in prostate cancer before and after neoadjuvant treatment with LH-RH analog, demonstrating that this therapy induced high AI and bax and survivin overexpression; thus acting as a proapoptotic agent in prostate cancer. A statistically significant correlation was found between high AI and overexpression of bax protein after therapy. It is probable that this kind of therapy is deeply implicated in promoting the apoptotic intrinsic pathway.

2003 - Disruption of beta1 integrin induces keratinocyte stem cell anoikis through the extrinsic apoptotic pathway [Abstract in Rivista]
Marconi, Alessandra; C., Fila; R., Tiberio; Giannetti, Alberto; Pincelli, Carlo
abstract

.

2003 - Expression and function of neurotrophins and their receptors in cultured human keratinocytes [Articolo su rivista]
Marconi, Alessandra; M., Terracina; C., Fila; J., Franchi; F., Bontè; G., Romagnoli; R., Maurelli; C. M., Failla; M., Dumas; Pincelli, Carlo
abstract

Whereas nerve growth factor has been extensively studied in human keratinocytes, little is known on the role of other members of the neurotrophin family. We investigated the expression and function of neurotrophins and neurotrophin receptors in cultured human keratinocytes. We demonstrated by reverse transcription-polymerase chain reaction that keratinocytes synthesize neurotrophin-3, brain-derived neurotrophic factor, and neurotrophin-4/5. These cells also express tyrosinase kinase A and C, the nerve growth factor and neuro-trophin-3 high-affinity receptors, respectively. On the other hand, only the truncated extracellular isoform of tyrosinase kinase B, the high-affinity brain-derived neurotrophic factor and neurotrophin-4/5 receptor, is detected in keratinocytes. Moreover, neurotrophin-3, brain-derived neurotrophic factor, and neurotrophin-4/5 proteins are secreted by human keratinocytes at low levels. Keratinocyte stem cells synthesize the highest amounts of nerve growth factor, while they secrete higher levels of nerve growth factor as compared with transit amplifying cells. Neurotrophin-3 stimulates keratinocyte proliferation, where brain-derived neurotrophic factor or neurotrophin-4/5 does not exert any effect on keratinocyte proliferation. Addition of neurotrophin-3 slightly upregulates the secretion of nerve growth factor, whereas nerve growth factor strongly augments neurotrophin-3 release. Ultraviolet B irradiation downregulates nerve growth factor, whereas it augments neurotrophin-3 and neurotrophin-4/5 protein levels. Ultraviolet A irradiation increases the level of neurotrophin-3, whereas it does not exert any effect on the other neurotrophins. Finally, neurotrophins other than nerve growth factor fail to protect human keratinocytes from ultraviolet B-induced apoptosis. This work delineates a functional neurotrophin network, which may contribute to epidermal homeostasis.

2003 - Expression and function of neurotrophins in human keratinocytes [Abstract in Rivista]
C., Fila; Marconi, Alessandra; M., Dumas; D., Baracchi; J., Franchi; F., Bonté; Pincelli, Carlo
abstract

.

2003 - Fas ligand in pemphigus sera induces keratinocyte apoptosis through the activation of caspase-8 [Articolo su rivista]
M., Puviani; Marconi, Alessandra; E., Cozzani; Pincelli, Carlo
abstract

The Fas/Fas ligand system triggers the extrinsic apoptotic pathway and is involved in several inflammatory conditions, also at the skin level. The Fas/Fas ligand cell death pathway plays a major role in anoikis, a type of apoptosis characterized by cell detachment. As pemphigus is characterized by loss of cell to cell adhesion, we evaluated the role of anoikis and Fas ligand in this bullous disease. We report that, in suprabasal epidermis from perilesional pemphigus skin, most keratinocytes are apoptotic. Moreover, Fas ligand levels are markedly increased in sera from pemphigus patients, whereas they are undetectable in sera from patients undergoing steroid treatment. Sera from untreated patients but not from patients under steroids induce keratinocyte apoptosis. Pemphigus-sera-induced cell death is partially inhibited by pretreatment with anti-Fas ligand antibodies and by incubation with caspase-8 inhibitor Z-IETD-FMK. Finally, caspase-8 is activated in keratinocytes provided with sera from pemphigus patients, whereas cleavage is partially blocked by pretreatment of sera with anti-Fas ligand antibody. These results suggest that increased Fas ligand in pemphigus sera is responsible for keratinocyte apoptosis, which occurs through the activation of a caspase-8-driven extrinsic apoptotic pathway.

2003 - The 'beauty' of skin neurobiology. [Articolo su rivista]
Pincelli, Carlo; F., Bontè
abstract

.

2003 - The p75 neurotrophin receptor modulates apoptosis and differentiation in a subpopulation of human keratinocytes [Abstract in Rivista]
D., Baracchi; Marconi, Alessandra; M., Pignatti; C., Fila; Pincelli, Carlo
abstract

.

2002 - Caspase-8-activated signal predominates in keratinocyte anoikis [Abstract in Rivista]
C., Fila; Marconi, Alessandra; S., Krajewski; J. C., Reed; Giannetti, Alberto; Pincelli, Carlo
abstract

.

2002 - Keratinocytes enriched for stem cells are protected from anoikis via an integrin signaling pathway in a Bcl-2 dependent manner [Articolo su rivista]
R., Tiberio; Marconi, Alessandra; C., Fila; C., Fumelli; M., Pignatti; S., Krajewski; Giannetti, Alberto; J. C., Reed; Pincelli, Carlo
abstract

Because inhibition of integrin signaling induces apoptosis, we investigated whether keratinocytes expressing beta1 and alpha6beta4 integrins (enriched for stem cells) are protected from cell death. Keratinocytes rapidly adhering to type IV collagen expressed highest levels of beta1 and alpha6beta4 and of the anti-apoptotic stem cell marker p63. Apoptotic cells were significantly higher in slowly adhering than in rapidly adhering keratinocytes. Anti-beta1 integrin caused a significant increase in apoptotic cells, while it decreased Bcl-2 levels in stem keratinocytes. Bax and Bad proteins were higher in slowly adhering than in rapidly adhering cells. By contrast, Bcl-2, Bcl-x and Mcl-1 proteins were highest in rapidly adhering keratinocytes and nearly absent in slowly adhering cells. After addition of anti-beta1 integrin, the apoptotic rate was significantly higher in HaCaT cells not expressing Bcl-2 than in controls. These results indicate that keratinocytes enriched for stem cells are protected from apoptosis via beta1 integrin, in a Bcl-2 dependent manner. (C) 2002 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.

2002 - Pemphigus sera, expressing high levels of Fas ligand, induces keratinocyte apoptosis [Abstract in Rivista]
Puviani, Mario; Marconi, Alessandra; Fila, Chiara; E., Cozzani; Giannetti, Alberto; Pincelli, Carlo
abstract

.

2002 - The low-affinity neurotrophin receptor p75 mediates the proapoptotic activity of brain-derived neurotrophic factor in keratinocytes [Abstract in Rivista]
Marconi, Alessandra; Fila, Chiara; M., Pignatti; Giannetti, Alberto; Pincelli, Carlo
abstract

.

2002 - The p75 neurotrophin receptor mediates spontaneous and UV-induced apoptosis in keratinocytes [Abstract in Rivista]
Marconi, Alessandra; M., Pignatti; Fila, Chiara; Puviani, Mario; Giannetti, Alberto; Pincelli, Carlo
abstract

.

2001 - Apoptosi e cute: dalla omeostasi epidermica alla cura del cancro. [Capitolo/Saggio]
Pincelli, Carlo
abstract

L'apoptosi è un processo fisiopatologico che interessa tutti i tessuti e le cellule del corpo umano. Tale processo serve all'organismo essenzialmente per liberarsi di cellule indesiderate. Nella cute un eccesso di apoptosi è coinvolto in malattie come il pemfigo, il lichen planus e il lupus, mentre un difetto di apoptosi è alla base di malattie iperproliferative come psoriasi e cancro

2001 - Innervazione Cutanea [Capitolo/Saggio]
Pincelli, Carlo; F., Fantini
abstract

La cute è l'organo più innervato del corpo umano. Il corpo cellulare del neurone risiede nelle radici dei gangli dorsali e le terminazioni nervose partono dalla cellula per raggiungere i tessuti periferici, tra cui la cute. Vi sono sia fibre nervose sensitive che del sistema autonomo. Le terminazioni nervose sono responsabili di raccogliere gli stimoli sensoriali e di trasmetterli al sistema nervoso. Inoltre le fibre simpatiche innervano vasi e ghiandole della cute

2001 - Stem keratinocytes are protected from cell death via an integrin signaling pathway, in a bcl-2-dependent manner [Abstract in Rivista]
R., Tiberio; Marconi, Alessandra; D., Ottani; S., Krajewski; J., Reed; Giannetti, Alberto; Pincelli, Carlo
abstract

.

2001 - The antiapoptotic bcl-2 proteins are highest and caspases are not activated in basal keratinocytes expressing high levels of p63 [Abstract in Rivista]
Marconi, Alessandra; C., Fila; S., Bertellini; M., Pignatti; S., Krajewski; J. C., Reed; Giannetti, Alberto; Pincelli, Carlo
abstract

.

2001 - The low-affinity neurotrophin receptor (p75) mediates apoptosis in human keratinocytes [Abstract in Rivista]
Marconi, Alessandra; C., Fila; S., Bertellini; M., Pignatti; Garuti, Cinzia; Giannetti, Alberto; Pincelli, Carlo
abstract

.

2000 - Autocrine nerve growth factor in human keratinocytes [Articolo su rivista]
Pincelli, Carlo; Marconi, Alessandra
abstract

Biologically active nerve growth factor (NGF) is synthesised and released by proliferating normal human keratinocytes. NGF up-regulates the expression of NGF mRNA in keratinocytes. Keratinocytes express both the low (p75)- and the high-affinity (TrkA) NGF-receptors, which are located in the basal layer of the epidermis. K252, a specific inhibitor of trk phosphorylation, blocks NGF-induced keratinocyte proliferation, in absence of exogenous NGF. Normal keratinocytes over-expressing TrkA proliferate better than control transfectants, while the NGF mimicking anti-Trk antibody induces an increased keratinocyte proliferation in Trk over-expressing cells as compared to mock transfected keratinocytes. In addition, NGF over-expressing keratinocytes proliferate better than mock transfected cells. K252, by blocking TrkA phosphorylation, induces apoptosis in normal keratinocytes, but not in keratinocytes over-expressing bcl-2. Further-more, NGF transfected keratinocytes are protected from UV-B-induced keratinocyte apoptosis, by maintaining constant levels of Bcl-2 and Bcl-x(L). Taken together these results support the concept of an autocrine survival system sustained by NGF and its high-affinity receptor in human keratinocytes. Because NGF and Trk levels are highly expressed in psoriasis, one could speculate that NGF autocrine system plays a role in the mechanisms associated with this and other hyperproliferative skin conditions, including cancer. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved.

2000 - C60 carboxyfullerene exerts a protective activity against oxidative stress-induced apoptosis in human peripheral blood mononuclear cells [Articolo su rivista]
D., Monti; L., Moretti; S., Salvioli; E., Straface; W., Malorni; R., Pellicciari; G., Schettini; M., Bisaglia; Pincelli, Carlo; C., Fumelli; M., Bonafe; C., Franceschi
abstract

C60 carboxyfullerene is a novel buckminsterfullerene-derived compound that behaves as a free-radical scavenger. In the present report, we investigated whether this drug exerts a protective activity against oxidative stress-induced apoptosis. Human peripheral blood mononuclear cells (PBMCs) were challenged by 2-deoxy-D-ribose (dRib) or TNF-alpha plus cycloheximide as agents that trigger apoptosis by interfering with the redox status of cell and mitochondrial membrane potential, We found that carboxyfullerene was able to protect quiescent PBMCs from apoptosis caused either by 8-deoxy-D-ribose or TNF-alpha plus cycloheximide by a mechanism partially involving the mitochondrial membrane potential integrity, known to be associated with early stages of apoptosis. These results represent the first indication for a target activity of buckminsterfullerenes on cells of the immune system and their mitochondria. (C) 2000 Academic Press.

2000 - Carboxyfullerenes protect human keratinocytes from ultraviolet-B-induced apoptosis [Articolo su rivista]
C., Fumelli; Marconi, Alessandra; S., Salvioli; E., Straface; W., Malorni; A. M., Offidani; R., Pellicciari; G., Schettini; Giannetti, Alberto; D., Monti; C., Franceschi; Pincelli, Carlo
abstract

Carboxyfullerene, a water-soluble carboxylic acid derivative of a fullerene, which acts as a free-radical scavenger, was investigated as a protective agent against ultraviolet-light-induced damage in human keratinocytes. First, we demonstrate that carboxyfullerene is not cytotoxic for these cells. In addition, this compound significantly reduces the ultraviolet-B-induced inhibition of keratinocyte proliferation and protects keratinocytes from apoptosis caused by ultraviolet B irradiation in a time- and dose-dependent fashion. Furthermore, the percentage of cells with depolarized mitochondria is significantly lower in ultraviolet-B-irradiated keratinocytes pretreated with carboxyfullerene than in cells provided with diluent alone. Carboxyfullerene also protects human keratinocytes from apoptosis induced by exposure to deoxy-D-ribose, a sugar that causes cell death through a pathway involving oxidative stress. On the other hand, ultraviolet B downregulates bcl-2 levels in human keratinocytes, and carboxyfullerene fails to prevent this effect. These results suggest that carboxy- fullerene protects human keratinocytes from ultraviolet B damage possibly via a mechanism interfering with the generation of reactive oxygen species from depolarized mitochondria without the involvement of bcl-2.

2000 - Keratinocyte nerve growth factor: more than just a neurotrophin [Relazione in Atti di Convegno]
Pincelli, Carlo; Marconi, Alessandra
abstract

.

2000 - Nerve growth factor and keratinocytes: a role in psoriasis [Articolo su rivista]
Pincelli, Carlo
abstract

Nerve growth factor (NGF) is synthesized and released by human keratinocytes. NGF acts as a neurotrophic molecule at the skin level, as it stimulates the sprouting of nerve fibers and regulates the synthesis and expression of neuropeptides. NGF can thus take part in neurogenic inflammation which in turn is involved in the pathogenesis of several inflammatory dermatoses. Human keratinocytes also synthesize and express the low (p75)-and the high-affinity (trk) NGF-receptor (NGF-R). NGF stimulates keratinocyte proliferation which is blocked by the natural alcaloid K252, a specific inhibitor of trk phosphorylation. K252 inhibits keratinocyte proliferation and induces keratinocyte apoptosis, in the absence of exogenous NGF, indicating the existence of an autocrine loop where NGF and trk act as key players. Finally, NGF protein levels are increased in psoriatic as compared to nonlesional and normal skin, and psoriatic keratinocytes express higher amounts of NGF than normal keratinocytes. This review will discuss the above findings in view of a possible involvement of NGF in the pathomechanisms associated with the development of the psoriatic lesion.

2000 - The free-radical scavengers carboxyfullerens protect human keratinocytes from UVB-induced apoptosis [Abstract in Rivista]
C., Fumelli; Marconi, Alessandra; Giannetti, Alberto; Pincelli, Carlo
abstract

.

1999 - C-3-Fullero-tris-methanodicarboxylic acid protects epithelial cells from radiation-induced anoikia by influencing cell adhesion ability [Articolo su rivista]
E., Straface; B., Natalini; D., Monti; C., Franceschi; G., Schettini; M., Bisaglia; C., Fumelli; Pincelli, Carlo; R., Pellicciari; W., Malorni
abstract

Anoikia is a type of apoptotic cell death that occurs in cells that are substrate-restricted in their growth. Buckminster-fullerenes represent a new class of chemical compounds with wide potential pharmacological antioxidant activity. In this report me provide the first demonstration that a water-soluble fullerene derivative, C-3-fullero-tris-methanodicarboxylic acid, synthesized in our laboratories, is capable of inducing anoikia resistance in Epithelial cells by a mechanism involving a 'trophic' effect on cell spreading-associated cytoskeletal components, i.e. on actin microfilaments. (C) 1999 Federation of European Biochemical Societies.

1999 - Expression of the novel inhibitor of apoptosis survivin in normal and neoplastic skin [Articolo su rivista]
C., Chiodino; A. M., Cesinaro; D., Ottani; F., Fantini; Giannetti, Alberto; G. P., Trentini; Pincelli, Carlo
abstract

Apoptosis plays a fundamental part in epidermal homeostasis, and apoptotic cells have been detected in normal and diseased skin. Little is known, however, on the inhibitory mechanisms of apoptosis at the skin level. In addition to bcl-2, a novel inhibitor of apoptosis designated survivin and structurally analogous to IAP apoptosis inhibitors has been recently identified. The expression of survivin in normal and pathologic skin was investigated. Immunohistochemical studies revealed that survivin is expressed in basal keratinocytes, but not in suprabasal epidermal layers, with a pattern similar to bcl-2, In western blots, the anti-survivin antibody recognized a single band of 16.5 kDa in protein extracts from normal human keratinocytes in culture, in agreement with the predicted size of survivin. In addition, survivin immunoreactivity was detected in benign and malignant melanocytic lesions, with strong expression in invasive lesions of melanomas. Whereas survivin staining was undetectable in benign epithelial tumors, such as seborrheic keratoses, it was observed in all epidermal layers in Bowen's disease. Interestingly, at variance with bcl-2, survivin was markedly expressed in squamous cell carcinoma, but virtually lacking in basal cell carcinoma, suggesting that these two apoptosis inhibitors may act through different anti-apoptotic pathways. Deregulation of survivin may influence both epidermal homeostasis and the development of melanoma and nonmelanoma skin cancer.

1999 - Keratinocytes that adhere most rapidly to type IV collagen and display characteristics of stem cells are protected from apoptosis [Abstract in Rivista]
Marconi, Alessandra; R., Tiberio; C., Fumelli; Giannetti, Alberto; Pincelli, Carlo
abstract

.

1999 - Nerve growth factor protects human keratinocytes from ultraviolet-B-induced apoptosis [Articolo su rivista]
Marconi, Alessandra; Vaschieri, Cristina; S., Zanoli; Giannetti, Alberto; Pincelli, Carlo
abstract

Ultraviolet radiation is a potent inducer of apoptosis, whereas autocrine nerve growth factor protects human keratinocytes from programmed cell death. To evaluate the role of nerve growth factor in the mechanisms of ultraviolet B-induced apoptosis, cultured human keratinocytes were ultraviolet B irradiated following pretreatment with K252, a specific inhibitor of the tyrosine kinase high-affinity nerve growth factor receptor. Here we report that the addition of K252 significantly enhanced keratinocyte apoptosis. We then transfected normal human keratinocytes with pNUT-hNGF, Nerve growth factor overexpressing keratinocytes secreted the highest amounts of nerve growth factor in culture supernatants, were more viable, and had a higher rate of proliferation than mock-transfected cells. Whereas ultraviolet B radiation downregulated nerve growth factor mRNA and protein as well as the tyrosine kinase high-affinity nerve growth factor receptor in normal keratinocytes, it failed to do so in nerve growth factor-transfected cells. Moreover, nerve growth factor overexpressing keratinocytes were partially resistant to apoptosis induced by increasing doses of ultraviolet B at 24 and 48 h. These results indicate that downregulation of nerve growth factor function plays an important part in the mechanisms of ultraviolet B-induced apoptosis in human keratinocytes. In addition, ultraviolet B caused a decrease in BCL-2 and BCL-x(L) expression in mock-transfected keratinocytes, but not in nerve growth factor overexpressing cells. Finally, nerve growth factor prevented the cleavage of the enzyme poly(ADP-ribose) polymerase induced in human keratinocytes by ultraviolet B, These results are consistent with a model whereby the autocrine nerve growth factor protects human keratinocytes from ultraviolet B-induced apoptosis by maintaining constant levels of BCL-2 and BCL-x(L), which in turn might block caspase activation.

1999 - The neurotrophin family in human keratinocytes [Abstract in Rivista]
Marconi, Alessandra; M., Terracina; C. M., Failla; G., Romagnoli; R., Maurelli; Pincelli, Carlo; Giannetti, Alberto; G., Zambruno
abstract

.

1998 - C2-ceramide triggers a BCL-2 mediated apoptosis in normal human keratinocytes. [Abstract in Rivista]
Di Nardo, A; Benassi, Luisa; Magnoni, Cristina; Pincelli, Carlo; Giannetti, Alberto
abstract

.

1998 - Carboxyfullerens reduce apoptotic death induced in cultured human keratinocytes by ultraviolet radiation. [Abstract in Rivista]
C., Fumelli; Marconi, Alessandra; A., Offidani; D., Monti; R., Pellicciari; Giannetti, Alberto; C., Franceschi; Pincelli, Carlo
abstract

.

1998 - Keratinocytes that adhere most rapidly to type IV collagen and express high levels beta 1 integrin do not undergo apoptosis. [Abstract in Rivista]
R., Tiberio; Marconi, Alessandra; Giannetti, Alberto; Pincelli, Carlo
abstract

.

1998 - Neuropeptides and Psoriasis. [Capitolo/Saggio]
Pincelli, Carlo
abstract

I cheratinociti sono le cellule maggiormente interessate dalla patologia psoriasica. Le alterazioni dei cheratinociti possono essere indotte dal rilascio di neuropeptidi nelle sedi lesionali. I neuropeptidi sono i protagonisti della infiammazione neurogena che è coinvolta nella psoriasi soprattutto quando questa è scatenata da eventi stressogeni. I livelli di nerve growth factor sono fortemente elevati nella psoriasi e la inibizione di questo fattore riduce la iperproliferazione cellulare, caratteristica fondamentale della patologia psoriasica.

1998 - Primary cutaneous nocardiosis [Articolo su rivista]
M., Folgaresi; G., Ferdani; Coppini, Maurizio; Pincelli, Carlo
abstract

A 37-year-old patient with systemic lupus erythematosus, who had been treated with oral corticosteroids for 10 years, developed primary cutaneous nocardiosis. Brown-violaceous, suppurative nodules and plaques arose on her right leg. Cultures of multiple biopsies on blood agar medium grew Nocardia. The patient received 500 mg of imipenem three times a day which resulted in complete regression of the lesions within two months. No relapse was observed 6 months later.

1998 - The role of nerve growth factor in ultraviolet radiation-induced cell cycle arrest and apoptosis of normal human keratinocytes. [Abstract in Rivista]
Marconi, Alessandra; Vaschieri, Cristina; Giannetti, Alberto; Pincelli, Carlo
abstract

.

1998 - Thymidine dimers induce a prolonged P53-dependent cell cycle arrest but not apoptosis in cultured human keratinocytes [Abstract in Atti di Convegno]
Magnoni, Cristina; Benassi, Luisa; A., Di Nardo; Bertazzoni, Giorgia; Giannetti, Alberto; Pincelli, Carlo
abstract

.

1997 - 1,25-dihydroxyvitamin D-3, transforming growth factor beta 1, calcium, and ultraviolet B radiation induce apoptosis in cultured human keratinocytes [Articolo su rivista]
Benassi, Luisa; D., Ottani; F., Fantini; Marconi, Alessandra; C., Chiodino; Giannetti, Alberto; Pincelli, Carlo
abstract

Apoptosis is a cellular process of self-directed suicide that plays a key role during morphogenesis and in the maintenance of homeostasis in continuously renewing tissues, Currently, apoptosis is detected mainly by gel electrophoresis of fragmented DNA and by typical ultrastructural features such as cell shrinkage and chromatin condensation, Recently, an in situ technique was developed that allows the detection of the apoptotic process in cells and the quantitation of apoptosis in cell populations, We applied this technique to evaluate the apoptotic process in cultured normal human keratinocytes under basic conditions and after stimulation with factors and agents that are presumed but have never been proved to induce apoptosis in these cells, Apoptosis was analyzed after stimulation with 1,25-dihydroxyvitamin D-3[1,25(OH)(2)D-3], transforming growth factor beta 1 (TGF beta 1), calcium, UVB, or tumor necrosis factor alpha (TNF alpha), All these factors except TNF alpha induced apoptosis in human keratinocytes. Whereas UVB and calcium were good apoptogenic stimuli at 6 and 24 h, respectively, the vitamin D derivative and TGF beta 1 induced apoptosis after 5 and 6 d in culture, Apoptosis was also established by DNA fragmentation and electron microscopy. Finally, TUNEL technique showed that the number of apoptotic cells increases slightly (5-10%) from 24 to 144 h even in untreated keratinocytes, Our studies indicate that factors normally involved in the regulation of cell growth and differentiation can also control apoptosis.

1997 - Acetyl-ceramide induced apoptosis in cultured human keratinocytes. [Abstract in Rivista]
Dinardo, A; Benassi, Luisa; Magnoni, Cristina; Pincelli, Carlo; Giannetti, Alberto
abstract

.

1997 - Autocrine nerve growth factor protects human keratinocytes from apoptosis through its high affinity receptor (TRK): A role for BCL-2 [Articolo su rivista]
Pincelli, Carlo; A. R., Haake; Benassi, Luisa; E., Grassilli; Magnoni, Cristina; D., Ottani; R., Polakowska; C., Franceschi; Giannetti, Alberto
abstract

Normal human keratinocytes synthesize and release nerve growth factor (NGF) and express both the low-and the high-affinity NGF receptor, Because NGF has been shown to rescue certain cell types from programmed cell death, we investigated the role of endogenous NGF in preventing keratinocyte apoptosis, We report here that apoptosis is induced in normal human keratinocytes in culture by blocking endogenous NGF signaling with either anti-NGF neutralizing antibody or K252, a specific inhibitor of the tyrosine kinase high-affinity NGF receptor. Apoptosis was assessed by DNA laddering, electron microscopy, and in situ nick end labeling technique, In anti-NGF-treated keratinocytes, the apoptotic process starts at 96 h, and is maximal at 120 h. After K252 treatment, apoptosis starts at 48 h and peaks at 120 h, Because the product of the bcl-2 proto-oncogene protects many cell types from apoptosis, we measured the levels of this protein in apoptotic keratinocytes, We found that both K252 and anti-NGF antibody strikingly downregulate bcl-2 expression, starting at 72 h. Furthermore, HaCat keratinocytes stably transfected with a plasmid containing bcl-2 cDNA fail to undergo apoptosis when treated with K252. These findings show that autocrine NGF acts as a survival factor for human keratinocytes in vitro through its high-affinity NGF receptor, possibly by maintaining constant levels of Bcl-2.

1997 - HaCat keratinocytes overexpressing bcl-2 are resistant to apoptosis induced by inhibition of autocrine nerve growth factor activity. [Abstract in Rivista]
Pincelli, Carlo; A., Haake; D., Ottani; Benassi, Luisa; Marconi, Alessandra; R., Polakowska; Giannetti, Alberto
abstract

.

1997 - Human keratinocytes overexpressing nerve growth factor are resistant to spontaneous and UV-induced apoptosis. [Abstract in Rivista]
Marconi, Alessandra; D., Ottani; Giannetti, Alberto; Pincelli, Carlo
abstract

.

1997 - Nerve growth factor and the skin [Articolo su rivista]
Pincelli, Carlo; M., Yaar
abstract

Nerve growth factor (NGF) is a neurotrophic polypeptide required for the survival and differentiation of sensory and sympathetic neurons (1, 2). NGF belongs to a small family of structurally and functionally related proteins termed neurotrophins (NTs), which include brain-derived neurotrophic factor (BDNF) (3), NT-3 (4) and NT-4/5 (5). All neurotrophic molecules exert their effect by binding two classes of transmembrane receptors. A low affinity receptor of similar to 75 kD (p75) (6), and a high affinity, tyrosine kinase receptor, of similar to 140 kD that belongs to the trk family of receptors (7, 8).

1997 - Neuropeptides, Nerve Growth Factor and Eczema [Capitolo/Saggio]
F., Fantini; Pincelli, Carlo; Giannetti, Alberto
abstract

L'eczema è una patologia infiammatoria della cute esacerbata dallo stress dove la neuroflogosi gioca un ruolo molto importante con up-regolazione dei livelli di neuropeptidi, NGF che modulano la attività di varie cellule cutanee

1997 - Role of cell-permeant ceramides in keratinocyte growth differentiation and apoptosis. [Abstract in Rivista]
A., Di Nardo; Benassi, Luisa; Pincelli, Carlo; Giannetti, Alberto
abstract

.

1997 - Thymidine dimers induce cell cycle arrest, but not apoptosis in cultured human keratinocytes. [Abstract in Rivista]
Magnoni, Cristina; Benassi, Luisa; Dinardo, A; Giannetti, Alberto; Pincelli, Carlo
abstract

.

1997 - Ultraviolet B-induced down-regulation of nerve growth factor and its high-affinity receptor (trk) mRNAs in human keratinocytes. [Abstract in Rivista]
Marconi, Alessandra; C., Chiodino; Vaschieri, Cristina; Giannetti, Alberto; Pincelli, Carlo
abstract

.

1997 - Ultraviolet radiations induce c-src activation in cultured human keratinocytes. [Abstract in Rivista]
Chiodino, C; Ottani, D; Magnoni, Cristina; Giannetti, Alberto; Pincelli, Carlo
abstract

.

1996 - Autocrine nerve growth factor protects human keratinocytes from apoptosis through its high-affinity receptor (trk): A role for BCL-2 [Abstract in Rivista]
Pincelli, Carlo; Magnoni, Cristina; Grassilli, E; Benassi, Luisa; Ottani, D; Franceschi, Claudio; Giannetti, Alberto
abstract

.

1996 - Il nerve growth factor endogeno protegge i cheratinociti umani dall'apoptosi attraverso il suo recettore ad alta affinità (trk):un ruolo per la bcl-2. [Abstract in Atti di Convegno]
D., Ottani; Magnoni, Cristina; E., Grassilli; Benassi, Luisa; Franceschi, Claudio; Giannetti, Alberto; Pincelli, Carlo
abstract

.

1996 - Neuropeptides, nerve growth factor and the skin [Articolo su rivista]
Pincelli, Carlo; F., Fantini; Giannetti, Alberto
abstract

Cutaneous nerve fibers can modulate inflammatory reactions through local release of neuropeptides (NP). In fact, NP are able to regulate both acute and chronic aspects of cutaneous inflammatory processes, such as vascular motility, cellular trafficking, activation and trophism. Several experimental evidences suggest a participation of NP to the pathogenesis of inflammatory dermatoses : in particular, modulation of inflammatory reactions in the skin can be obtained through pharmacologic manipulation of the neuropeptidergic system. Furthermore, evidence of local alterations of NP levels in inflamed skin has been obtained. It has been hypothesized that the inflammatory process induces reactive changes in the neuronal NP content. The primary candidate as mediator of this neuronal recruitment in the course of inflammatory reactions is the neurothrophin nerve growth factor, that is produced in the skin in increased amounts during inflammatory processes and may regulate peptidergic synthesis at a neuronal level.

1996 - Transforming growth factor-beta, 1,25-dihydroxyvitamin D3, calcium and ultraviolet B radiation, but not tumor necrosis factor-alpha induce apoptosis in cultured human keratinocytes. [Abstract in Rivista]
Benassi, Luisa; D., Ottani; F., Fantini; Giannetti, Alberto; Pincelli, Carlo
abstract

.

1996 - Ultraviolet B radiations down regulate nerve growth factor mRNA and release in human keratinocytes. [Abstract in Rivista]
Marconi, Alessandra; C., Chiodino; Vaschieri, Cristina; Giannetti, Alberto; Pincelli, Carlo
abstract

.

1995 - A Dual Role For P75 Nerve Growth-Factor Receptor In Melanocyte Survival And Apoptosis [Abstract in Rivista]
M., Yaar; S., Zhai; Pincelli, Carlo; B. A., Gilchrest
abstract

.

1995 - Expression and function of nerve growth factor (NGF) and NGF receptor on keratinocytes. [Abstract in Atti di Convegno]
Pincelli, Carlo; Magnoni, Cristina; M., Yaar; B. A., Gilchrest; Giannetti, Alberto
abstract

.

1995 - K252a, A High-Affinity NGF-Receptor (Trk) Inhibitor, Induces Apoptosis In Human Keratinocytes [Abstract in Rivista]
Pincelli, Carlo; Magnoni, Cristina; E., Grassilli; Benassi, Luisa; D., Ottani; C., Franceschi; Giannetti, Alberto
abstract

.

1995 - K252a, a high affinity NGF-receptor (trk) inhibitor, induces apoptosis in cultured human keratinocytes. [Abstract in Atti di Convegno]
Magnoni, Cristina; E., Grassilli; Benassi, Luisa; D., Ottani; Franceschi, Claudio; Pincelli, Carlo; Giannetti, Alberto
abstract

.

1995 - Nerve Growth-Factor Is Increased In Psoriatic Skin [Articolo su rivista]
F., Fantini; Magnoni, Cristina; L., Bracci Laudiero; Pincelli, Carlo
abstract

.

1995 - Nerve Growth-Factor Receptor And Neurochemical Markers In Human Oral-Mucosa - An Immunohistochemical Study [Articolo su rivista]
F., Fantini; Giannetti, Alberto; Benassi, Luisa; V., Cattaneo; Magnoni, Cristina; Pincelli, Carlo
abstract

Background: The innervation of the oral mucosa has so far been studied mainly by histochemical and ultrastructural techniques. Only few studies have investigated the presence of neural proteins and neurotransmitters in human gingival mucosa. Objective: The purpose of the present study was to evaluate the presence and distribution of neural structural and transmitter proteins in different areas of normal human oral mucosa. Method: Indirect immunofluorescence was employed on specimens taken from different mucosal regions (gingiva, lips, gums, palate). Both structural (low-affinity nerve growth factor receptor, NGFr; protein gene product 9.5, PGP 9.5) and neuropeptide markers (substance P; calcitonin gene-related peptide; vasoactive intestinal peptide, neuropeptide Y) were used. Results: NGFr and PGP 9.5 intensely labelled both nerve fibres and selected epithelial cells, while neuropeptide immunoreactivity was scarcely expressed and exclusively localized in nerve fibres, Conclusion: Similarity in the distribution pattern and neurochemistry between oral and cutaneous innervation is apparent. Expression of NGFr could be relevant to the trophism of both the oral innervation and epithelium.

1995 - Neurogenic Inflammation And The Skin Neural Modulation Of Cutaneous Inflammatory Reactions [Articolo su rivista]
F., Fantini; Magnoni, Cristina; Pincelli, Carlo; Giannetti, Alberto
abstract

This review presents current concepts in neurogenic inflammation, with particular regard to the skin. Although the participation of the peripheral nervous system in the development of inflammatory reactions has been known for a long time, recent advances in the understanding of the distribution, functions and targets of neural mediators has greatly enhanced interest and research on this topic. A role for peripheral nerve terminals not only in the vascular responses, but also in the modulation of inflammatory cell activities, has been proposed. Neuropeptides released from nerve endings could intervene in the various phases of the inflammatory process, contributing to the complex network of inflammatory mediators which locally regulate cell function, trafficking and trophism. An increasing body of evidence also supports an important role for these fibers in the pathogenesis of both spontaneous and experimentally-induced cutaneous inflammatory lesions. Indeed, recent findings indicate that the different neural transmitters take part in the mechanisms of cutaneous inflammatory reactions with specific, differentiated and often protective effects. Therefore, we would suggest that this involvement could be more properly described as neurogenic modulation of inflammatory reactions.

1995 - The Role Of Nerve Growth-Factor In Preventing Keratinocyte Apoptosis [Abstract in Rivista]
S., Zhai; Pincelli, Carlo; M., Yaar; J., Gonsalves; B. A., Gilchrest
abstract

.

1994 - Expression And Function Of Nerve Growth-Factor And Nerve Growth-Factor Receptor On Cultured Keratinocytes. [Articolo su rivista]
Pincelli, Carlo; C., Sevignani; Manfredini, Rossella; Grande, Alexis; F., Fantini; L., Bracci Laudiero; L., Aloe; Ferrari, Sergio; Cossarizza, Andrea; Giannetti, Alberto
abstract

Keratinocytes, a key cellular component both for homeostasis and pathophysiologic processes of the skin, secrete a number of cytokines and are stimulated by several growth factors. Nerve growth factor (NGF) is synthesized in the skin and basal keratinocytes express the low-affinity nerve growth factor receptor (NGF-R). We present evidence that normal human keratinocytes in culture express the low- and the high-affinity NGF-R both at the mRNA level, as determined by reverse-transcription polymerase chain reaction and at the protein level, as shown by cytofluorimetric analysis. NGF significantly stimulates the proliferation of normal human keratinocytes in culture in a dose-dependent manner. This effect can be prevented by the addition of both an anti-NGF neutralizing antibody and a high-affinity NGF-R (trk) specific inhibitor, the natural alkaloid K252a. By contrast, keratinocyte proliferation is not inhibited by an anti - low-affinity NGF-R monoclonal antibody, thus suggesting that NGF effect on human keratinocytes is mediated by the high-affinity NGF-R. Moreover, NGF mRNA is expressed in normal human keratinocytes and NGF is secreted by keratinocytes in increasing amounts during growth, as detected by enzyme-linked immunosorbent assay. These results suggest that NGF could act as a cytokine in human skin and take part in disorders of keratinocyte proliferation.

1994 - Il nerve growth factor è aumentato nella cute psoriasica. [Abstract in Atti di Convegno]
Pincelli, Carlo; F., Fantini; Magnoni, Cristina; E., Distratis; Benassi, Luisa; L., Aloe; Giannetti, Alberto
abstract

.

1994 - Il nerve growth factor, ma non la neurotrofina-3 stimola la proliferazione dei cheratinociti umani in coltura. [Abstract in Atti di Convegno]
Magnoni, Cristina; Benassi, Luisa; F., Fantini; Sevignani, C.; Giannetti, Alberto; Pincelli, Carlo
abstract

.

1994 - Nerve Growth-Factor And The Skin [Articolo su rivista]
Pincelli, Carlo; F., Fantini; Giannetti, Alberto
abstract

.

1994 - Neural Markers And Cutaneous Inflammation [Articolo su rivista]
Pincelli, Carlo; F., Fantini
abstract

.

1994 - Notalgia-Paresthetica - Clinical, Pathological And Immunohistochemical Observations In 12 Cases [Articolo su rivista]
F., Fantini; F., Zorzi; G., Rizzitelli; Benassi, Luisa; Pincelli, Carlo
abstract

Notalgia paresthetica (NP) is a perculiar dermatosis characterized by pigmented and pruritic patches on the skin of the back. Although not a rare condition, it has been infrequently reported in the literature. Recent immunohistochemical studies, as well as the effectiveness of topical capsaicin in relieving NP symptoms, point to a pathogenetic role of cutaneous nerve fibers in NP. We present here our clinical and pathological observations in 12 cases of NP. In eight cases skin specimens were also immunocytochemically evaluated with a panel of neurochemical markers, and compared to both the contralateral skin of the dorsum and the site-matched skin from healthy subjects. No significant difference was observed in the presence, distribution or density of immunoreactive fibers in the different groups. The results suggest that functional, rather than morphological changes in local nerve terminals could be operating in this disease.

1994 - Psoriasis And The Nervous-System [Articolo su rivista]
Pincelli, Carlo; F., Fantini; Magnoni, Cristina; Giannetti, Alberto
abstract

Both clinical and experimental evidence is accumulating on the role of the nervous system in the pathogenesis of psoriasis. Sporadic reports as well as extensive studies indicate that emotional stress can act as an exacerbating event in psoriasis. Moreover, that neurogenic mechanisms are operating in psoriasis is suggested by clinical, pharmacologic and experimental data. We have focused our investigations on the role of vasoactive intestinal peptide (VIP) and substance P (SP) in psoratic lesions using a variety of experimental approaches: 1) receptor autoradiography; 2) immunohistochemistry; 3) radio-imunoassay; 4) human keratinocytes cultures. Our results indicate that an imbalance of VLP and SP exists in psoriatic lesions. and that these neuropeptides exert different and specific effects on human keratinocytes. At present, however, the finding of psoriasis being exacerbated by psychological factors cannot be satisfactorily explained merely by alterations of neuropeptides in the skin.

1994 - Psoriasis and nervous system [Capitolo/Saggio]
Pincelli, Carlo; F., Fantini; Giannetti, Alberto
abstract

Nella cute psoriasica la infiammazione neurogena è regolata dalle terminazioni nervose che rilasciano neuropeptidi. La sostanza P per esempio è diminuita mentre il VIP è aumentato nella cute lesionale. Questi meccanismi contribuiscono allo scatenamento della malattia psoriasica soprattutto in situazioni di stress psicologico

1993 - Autoradiographic detection of substance P receptors in normal and psoriatic skin [Articolo su rivista]
Pincelli, Carlo; F., Fantini; L., Giardino; M., Zanni; L., Calzà; C., Sevignani; Giannetti, Alberto
abstract

Substance P has been detected in human skin and has been implicated in the pathogenesis of certain inflammatory cutaneous disorders. However, little is known about the number and distribution of substance P binding sites in the skin. Receptor autoradiography was employed to detect and quantitate substance P receptors in normal as well as psoriatic skin. Substance P binding sites were distributed in the epidermis and dermis both in normal and psoriatic skin. In the dermis, the highest densities of SP binding sites were found in the areas corresponding to the dermal papillae and the adnexal structures. Quantitative analysis revealed that saturable binding was obtained both in the epidermis and in the labeled dermal areas. Rosenthal plot values were consistent with a single population of binding sites. No difference in the binding measurements was observed between normal and psoriatic skin. The presence of substance P receptors in the epidermis and in the dermal papillae raises interesting issues on the possible targets of this peptide in human skin both under physiologic and pathologic conditions.

1993 - Neuropeptides And Skin Inflammation [Articolo su rivista]
Pincelli, Carlo; F., Fantini; Giannetti, Alberto
abstract

Neuropeptides (NP) are protein compounds contained both in the central and peripheral nervous system. They can be antidromically released from sensory nerves and are implicated in the so-called neurogenic inflammation. They also exert a number of functions within the immune system and are thought to act as trophic as well as mitogenic substances. Several NP have been detected in human skin by immunohistochemical and radioimmunological techniques, and recent reports have demonstrated that NP could be involved in the mechanisms of certain dermatoses. The involvement of NP in either physiological or pathophysiological skin conditions is discussed. Moreover, a few questions, which still need to be addressed, are raised, and future directions this field of research should take are outlined.

1993 - Normal Human Keratinocytes Express Nerve Growth-Factor Receptor And Proliferate In Response To Nerve Growth-Factor [Abstract in Rivista]
Pincelli, Carlo; C., Sevignani; Grande, Alexis; F., Fantini; L., Aloe; Giannetti, Alberto
abstract

.

1993 - Pigmented Contact-Dermatitis From Deodorant [Articolo su rivista]
Pincelli, Carlo; R., Magni; A., Motolese
abstract

.

1992 - Cutaneous Innervation In Chronic-Renal-Failure Patients - An Immunohistochemical Study [Articolo su rivista]
F., Fantini; A., Baraldi; C., Sevignani; A., Spattini; Pincelli, Carlo; Giannetti, Alberto
abstract

Most chronic renal failure patients suffer from generalized pruritus. An involvement of cutaneous nerve terminals in the pathogenesis of uremic pruritus has been suggested. Skin specimens from 24 uremic patients and 10 healthy subjects were processed with an indirect immunofluorescence method to investigate the presence and distribution of a number of neuronal markers and neuropeptides. No difference was found between the two groups in the distribution pattern of the positive nerve fibres. However, a reduction in the total number of skin nerve terminals in the uremic patients was detected. No correlation could be found between the immunohistochemical findings and the clinical features. Our results suggest that the skin innervation is altered in most chronic renal failure patients, possibly as a consequence of neuropathy.

1992 - Neuropeptidi e cute: il peptide vasoattivo intestinale nelle dermatosi infiammatorie. [Articolo su rivista]
Pincelli, Carlo; Magnoni, Cristina; F., Fantini; Giannetti, Alberto
abstract

.

1992 - Substance P is diminished and vasoactive intestinal peptide is augmented in psoriatic lesions and these peptides exert disparate effects on the proliferation of cultured human keratinocytes [Articolo su rivista]
Pincelli, Carlo; F., Fantini; P., Romualdi; C., Sevignani; G., Lesa; Benassi, Luisa; Giannetti, Alberto
abstract

An involvement of neurogenic components in the pathogenesis of psoriatic lesions has been suggested and neuropeptides are thought to play a modulatory role in cutaneous inflammation. In this study, we evaluated the immunoreactivity of the neuropeptides vasoactive intestinal polypeptide (VIP) and substance P (SP) in the skin of patients with chronic plaque psoriasis, by immunohistochemistry and radioimmunoassay. No differences were observed, by immunohistochemistry, in the expression and localization of VIP and SP between psoriatic and normal skin. Using the radioimmunologic technique on whole skin homogenates, VIP levels were significantly increased in psoriatic lesions as compared to normal skin. By contrast, SP levels were significantly lower in lesional and non-lesional psoriatic skin than in normal skin. In addition, we examined the effect of VIP and SP on the proliferation of cultured normal human keratinocytes. VIP (1-28) (1 nM-1 microM) as well as VIP fragments (10-28) (1 nM-1 microM) and (22-28) (1 nM-1 microM) stimulated the proliferation of keratinocytes in a dose-dependent manner, whereas the VIP fragment (1-12) (1 nM-1 microM) was ineffective. The VIP antagonist (N-Ac-Tyr1, D-Phe2)-GRF (1-29)-NH2 (0.1 microM) significantly inhibited the VIP effect on keratinocytes. On the other hand, SP (0.1 microM) not only failed to stimulate keratinocyte growth, but also blocked the VIP-induced stimulation of these cells. The imbalance of cutaneous VIP and SP and their disparate effects on the proliferation of normal human keratinocytes in culture would suggest that these peptides are involved in the pathogenesis of psoriasis and may exert different modulatory activities in the mechanisms underlying the psoriatic lesion

1992 - Vasoactive Intestinal Polypeptide And Substance-P In The Pathogenesis Of Atopic-Dermatitis [Articolo su rivista]
Giannetti, Alberto; F., Fantini; A., Cimitan; Pincelli, Carlo
abstract

Neurogenic components are probably involved in the pathogenesis of atopic dermatitis (AD) and several neuropeptides have been implicated in the mechanisms underlying this disease. The aim of the present study was to evaluate by radio-immunoassay (RIA), the vasoactive intestinal polypeptide (VIP) and substance P (SP) content in whole-skin homogenates of AD lesions. RIA was performed using an antiserum, AH78, recognizing the carboxy-terminal fragment VIP (22-28) and a polyclonal antiserum directed against SP. VIP levels were markedly increased in lesional AD skin (5.62 +/- 1.25 pmol/g tissue) vis-avis controls (0.43 +/- 0.08 pmol/g tissue), whereas SP levels were significantly lower in lesional skin (0.25 +/- 0.03 pmol/g tissue) than in normal skin (0.97 +/- 0.24 pmol/g tissue). The results confirm that VIP and SP are relevant to the pathogenesis of AD and their imbalance might reflect diverse roles of these NP in the modulation of AD lesion.

1991 - Langerhans Cells Can Express Neuron-Specific Enolase Immunoreactivity [Articolo su rivista]
F., Fantini; Pincelli, Carlo; C., Sevignani; A., Baraldi; Giannetti, Alberto
abstract

.

1991 - Neuron-Specific Enolase Is A Marker Of Cutaneous Langerhans Cell Histiocytosis (X)-A Comparative-Study With S100 Protein [Articolo su rivista]
J., Kanitakis; F., Fantini; Pincelli, Carlo; C., Hermier; D., Schmitt; J., Thivolet
abstract

The immunohistochemical expression of neuron-specific enolase (gamma/gamma) (NSE) was studied comparatively with S100 protein in a group of Langerhans-cell-type ("X") (n = 8) and non-Langerhans-cell-type ("non X") (n = 24) cutaneous histiocytoses. NSE was expressed by the majority (70-90%) of histiocytic cells in all cases of Langerhans-cell histiocytoses, whereas it was absent from non-Langerhans-cell histiocytoses. S100 protein was expressed by the majority of Langerhans-cell histiocytosis cells but also by a small percentage (1-5%) of cells in non Langerhans-cell histiocytoses. These results show that NSE is almost as sensitive as, but more specific than, S100 protein in discriminating Langerhans-cell from non-Langerhans cell cutaneous histiocytoses, and that it consequently represents a useful adjunct in the immunohistochemical diagnosis of histiocytic skin diseases.

1991 - Skin Levels Of Vasoactive Intestinal Polypeptide In Atopic-Dermatitis [Articolo su rivista]
Pincelli, Carlo; F., Fantini; P., Romualdi; G., Lesa; Giannetti, Alberto
abstract

Atopic dermatitis (AD) can be exacerbated by various factors, including emotional stress, scratching and sweating. The aim of the present study was to evaluate the hypothesis that the inflammatory reaction in AD is also neurogenic. For this purpose, the levels of vasoactive intestinal polypeptide were measured radioimmunologically in whole-tissue homogenates of lesional skin of 13 patients with atopic dermatitis. Radioimmunoassay was performed using an antiserum, AH78, recognizing the carboxy-terminal fragment vasoactive intestinal polypeptide (22-28). Vasoactive intestinal polypeptide immunoreactivity was detected in relatively low amounts in control skin (0.428 +/- 0.08 pmol/g tissue), whereas a marked increase in the peptide was observed in lesional skin of patients with atopic dermatitis (5.62 +/- 1.25 pmol/g tissue). These results seem to suggest that vasoactive intestinal polypeptide could have a pathogenetic relevance in skin lesions of atopic dermatitis.

1990 - Erythema multiforme associated with contact sensitization. Description of 6 clinical cases [Articolo su rivista]
Seidenari, Stefania; A., Di Nardo; A., Motolese; Pincelli, Carlo
abstract

Allergic contact dermatitis generally presents itself as an eczematous eruption. However, some allergens, such as plants, exotic woods, topical medications and chemicals have been reported as causing an erythema multiforme-like eruption. We describe 6 patients, 4 women and 2 men ranging from 28 to 66 years. They showed erythematous urticarial papules, plaques and target-lesions similar to the manifestations previously reported as "erythema multiforme-like eruption". On the basis of the positive epicutaneous tests, the aetiological agents of these reactions are presumed to be Idoxuridine, textile dyes, deodorants, paraphenylenediamine and pyrrolnitrin. The clinical pattern, the histopathological features and the pathogenesis are discussed. Finally, the clinical and anamnestic criteria indicating the opportunity of performing patch tests in patients showing an atypical erythema multiforme are suggested.

1990 - Local-Effects Of Granulomatous Inflammation On Functional Activation Of T-Cells In Athymic Mice [Articolo su rivista]
A., Fujioka; H., Suya; Pincelli, Carlo; A., Hashimoto; K., Fukuyama; W. L., Epstein
abstract

.

1990 - Neuron-Specific Enolase-Immunoreactive Fibers In Uremic Patients [Articolo su rivista]
F., Fantini; A., Baraldi; Pincelli, Carlo
abstract

.

1990 - Neuropeptide Y-Like Immunoreactivity In Langerhans Cells From Patients With Atopic-Dermatitis [Articolo su rivista]
Pincelli, Carlo; F., Fantini; P., Massimi; Giannetti, Alberto
abstract

.

1990 - Neuropeptide-Like Immunoreactivity In Skin-Lesions Of Atopic-Dermatitis And Psoriasis [Articolo su rivista]
F., Fantini; Pincelli, Carlo; P., Massimi; Giannetti, Alberto
abstract

.

1990 - Neuropeptides In Skin From Patients With Atopic-Dermatitis - An Immunohistochemical Study [Articolo su rivista]
Pincelli, Carlo; F., Fantini; P., Massimi; G., Girolomoni; Seidenari, Stefania; Giannetti, Alberto
abstract

.

1990 - Relationship Between Nk Cells And Granulomatous Inflammation In Mice [Articolo su rivista]
A., Hashimoto; Pincelli, Carlo; A., Fujioka; K., Fukuyama; W. L., Epstein
abstract

The relationship between natural killer (NK) cells and granulomatous inflammation was investigated using two experimental granuloma models in C57BL/6 mice. The hepatic granuloma model was produced by infection with Schistosoma mansoni (S. mansoni), and the skin granuloma model was established by subcutaneous inoculation of the isolated hepatic granulomas. NK cell activity in lymph nodes and spleen, measured by Cr-51-release assay against YAC-1 cells, was compared to that in age-matched control mice. The activity decreased progressively as granulomas developed without changing the number of NK-1.1+ cells. The reduced NK cell activity was not reversed by administration of indomethacin. In order to further substantiate whether NK cells contribute to granulomatous inflammation, NK cells were depleted in the mice by injection of anti-NK-1.1 mAb. Reduction of NK cell activity (70-90%) was achieved during granuloma formation. An increase of about 20% in the mean granuloma diameter was detected in the mAb-treated mice in both models. Moreover, the percentage of granuloma takes in the skin model was enhanced (65% increase). The mAb treatment did not alter T cell counts in granulomas, T cell subset numbers or proliferative response in spleen. These findings indicate that NK cells directly play a regulatory role in granulomatous inflammation.

1989 - Immunosuppression And In vivo T-Cell Function In Granulomatous Inflammation [Articolo su rivista]
A., Fujioka; Pincelli, Carlo; A., Hashimoto; K., Fukuyama; W. L., Epstein
abstract

.

1988 - Immunopathologic Studies In Pityriasis Lichenoides [Articolo su rivista]
Giannetti, Alberto; G., Girolomoni; Pincelli, Carlo; Benassi, Luisa
abstract

.

1988 - Skin Granuloma-Formation In Mice Immunosuppressed By Cyclosporine [Articolo su rivista]
H., Suya; A., Fujioka; Pincelli, Carlo; K., Fukuyama; W. L., Epstein
abstract

.

1988 - T-Cell Depletion By Monoclonal-Antibodies Does Not Prevent Granuloma-Formation In Mice [Articolo su rivista]
PINCELLI, Carlo; A., Fujioka; A., Hashimoto; K., Fukuyama; W. L., Epstein
abstract

.

1987 - Pseudopelade Of Brocq - An Immunologically Mediated Disease [Articolo su rivista]
Pincelli, Carlo; G., Girolomoni; Benassi, Luisa
abstract

.

1986 - Effect Of Selective Enzymatic Digestions On Skin Biopsies From Pseudoxanthoma Elasticum - An Ultrastructural-Study [Articolo su rivista]
Contri, Miranda; Pincelli, Carlo; Bertazzoni, G. M.; Ronchetti, Ivonne
abstract

.

1986 - Expression Of T6-Antigen On The Epidermal-Keratinocytes In Various Dermatoses .2. [Articolo su rivista]
Pincelli, Carlo; G., Girolomoni; Benassi, Luisa
abstract

.

1986 - Phenytoin-Treatment Of Rdeb Vegetans [Articolo su rivista]
Pini, Luigi Alberto; G., Girolomoni; Coppini, Maurizio; Pincelli, Carlo
abstract

To assess the potential short- and long-term efficacy of phenytoin in treating recessive dystrophic epidermolysis bullosa (RDEB), we treated a patiens with therapeutic doses of oral phenytoin (blood level, 8 to 10 micrograms/mL) for an 10 moths period During this tretament we recorded a percentage decrease in blistering of the skin over 50% This report suggest that phenytoin seems to have therapeutic efficacy in RDEB.

1985 - Urinary techniques of glycosaminoglycans, hydroxyproline and L-glutamic acid in psoriatic patiens [Articolo su rivista]
Plessi, Maria; Parenti, Carlo; Monzani, Agar; Coppini, Dino; Pincelli, Carlo; G., Vaccari
abstract

Urinary glycosaminoglycans and hydroxyproline were spectrophotometrically determined in a series of psoriatic patients. In addition, urinary L-glutamic acid was evaluated by an enzymatic technique. The results demontrated that the psoriatic patients have considerably higher concentrations of all three metabolites than healty subjects.

1984 - Definition of health education. Current status of Italian legislation. Structures foreseen by the law [Articolo su rivista]
Coppini, Maurizio; Pincelli, Carlo
abstract

.

1982 - Psoriasis and lithium carbonate. Apropos of 2 clinical cases [Articolo su rivista]
Pincelli, Carlo; Coppini, Maurizio; M., Zucchi; M., Zanini
abstract

.

1980 - The Absence Of Sezary Cells In The Peripheral-Blood Of Patients With Generalized Inflammatory Dermatoses [Articolo su rivista]
Pincelli, Carlo; A. J., Dean; A. P., Warin
abstract

.