Effects of CTLA4-Ig treatment on circulating fibrocytes and skin fibroblasts from the same systemic sclerosis patients: an in vitro assay

Abstract Background Systemic sclerosis (SSc) is characterized by vasculopathy and progressive fibrosis. CTLA4-Ig (abatacept) is able to interact with the cell surface costimulatory molecule CD86 and downregulate the target cell. The aim of this study was to evaluate the in-vitro effects of CTLA4-Ig...

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Main Authors:	Maurizio Cutolo, Stefano Soldano, Paola Montagna, Amelia Chiara Trombetta, Paola Contini, Barbara Ruaro, Alberto Sulli, Stefano Scabini, Emanuela Stratta, Sabrina Paolino, Carmen Pizzorni, Vanessa Smith, Renata Brizzolara
Format:	Article
Language:	English
Published:	BioMed Central 2018-07-01
Series:	Arthritis Research & Therapy
Subjects:	Fibrocytes Skin fibroblasts CTLA4-Ig Systemic sclerosis Connective tissue disease
Online Access:	http://link.springer.com/article/10.1186/s13075-018-1652-6

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spelling	doaj-art-8fb2d27c8780487180edfa49ae8eb1c22018-08-20T18:27:42ZengBioMed CentralArthritis Research & Therapy1478-63622018-07-012011910.1186/s13075-018-1652-6Effects of CTLA4-Ig treatment on circulating fibrocytes and skin fibroblasts from the same systemic sclerosis patients: an in vitro assayMaurizio Cutolo0Stefano Soldano1Paola Montagna2Amelia Chiara Trombetta3Paola Contini4Barbara Ruaro5Alberto Sulli6Stefano Scabini7Emanuela Stratta8Sabrina Paolino9Carmen Pizzorni10Vanessa Smith11Renata Brizzolara12Research Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genoa, IRCCS San Martino Polyclinic HospitalResearch Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genoa, IRCCS San Martino Polyclinic HospitalResearch Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genoa, IRCCS San Martino Polyclinic HospitalResearch Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genoa, IRCCS San Martino Polyclinic HospitalDivision of Clinical Immunology, Department of Internal Medicine, University of GenoaResearch Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genoa, IRCCS San Martino Polyclinic HospitalResearch Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genoa, IRCCS San Martino Polyclinic HospitalOncologic Surgery, Department of Surgery, IRCCS San Martino PolyclinicOncologic Surgery, Department of Surgery, IRCCS San Martino PolyclinicResearch Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genoa, IRCCS San Martino Polyclinic HospitalResearch Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genoa, IRCCS San Martino Polyclinic HospitalDepartment of Rheumatology, Ghent University Hospital, Ghent UniversityResearch Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genoa, IRCCS San Martino Polyclinic HospitalAbstract Background Systemic sclerosis (SSc) is characterized by vasculopathy and progressive fibrosis. CTLA4-Ig (abatacept) is able to interact with the cell surface costimulatory molecule CD86 and downregulate the target cell. The aim of this study was to evaluate the in-vitro effects of CTLA4-Ig treatment on circulating fibrocytes and skin fibroblasts isolated from the same SSc patient. Methods Circulating fibrocytes and skin fibroblasts were obtained from eight SSc patients with “limited” cutaneous involvement and from four healthy subjects (HSs). Samples were analyzed by fluorescence-activated cell sorter analysis (FACS) at baseline (T0) and after 8 days of culture (T8) for CD45, collagen type I (COL I), CXCR4, CD14, CD86, and HLA-DRII expression. Circulating fibrocytes were treated for 3 h and skin fibroblasts for 24/48 h with CTLA4-Ig (10, 50, 100, 500 μg/ml). Quantitative real-time polymerase chain reaction (qRT-PCR) was performed for CD86, COL I, FN, TGFβ, αSMA, S100A4, CXCR2, CXCR4, CD11a, and Western blotting was performed for COL I and FN. Results Using qRT-PCR, the T8-cultured SSc circulating fibrocytes which had not been treated with CTLA4-Ig showed higher gene expression for CD86, αSMA, S100A4, TGFβ, and COL I compared with HS circulating fibrocytes. Interestingly, αSMA/COL I gene expression was significantly lower only in the SSc circulating fibrocytes treated with CTLA4-Ig for 3 h (p < 0.01, p < 0.05). On the contrary, no effects were observed for either SSc or HS skin fibroblasts after CTLA4-Ig treatment. COL I and FN protein expression was unchanged in both SSc and HS skin fibroblasts by Western blot. Conclusions Circulating fibrocytes seem to be more responsive to CTLA4-Ig treatment than skin fibroblasts from the same SSc patient, likely due to their higher expression of CD86. CTLA4-Ig treatment might downregulate the fibrotic process in SSc patients by downregulating the fibrocytes, circulating progenitor cells.http://link.springer.com/article/10.1186/s13075-018-1652-6FibrocytesSkin fibroblastsCTLA4-IgSystemic sclerosisConnective tissue disease
institution	Open Data Bank
collection	Open Access Journals
building	Directory of Open Access Journals
language	English
format	Article
author	Maurizio Cutolo Stefano Soldano Paola Montagna Amelia Chiara Trombetta Paola Contini Barbara Ruaro Alberto Sulli Stefano Scabini Emanuela Stratta Sabrina Paolino Carmen Pizzorni Vanessa Smith Renata Brizzolara
spellingShingle	Maurizio Cutolo Stefano Soldano Paola Montagna Amelia Chiara Trombetta Paola Contini Barbara Ruaro Alberto Sulli Stefano Scabini Emanuela Stratta Sabrina Paolino Carmen Pizzorni Vanessa Smith Renata Brizzolara Effects of CTLA4-Ig treatment on circulating fibrocytes and skin fibroblasts from the same systemic sclerosis patients: an in vitro assay Arthritis Research & Therapy Fibrocytes Skin fibroblasts CTLA4-Ig Systemic sclerosis Connective tissue disease
author_facet	Maurizio Cutolo Stefano Soldano Paola Montagna Amelia Chiara Trombetta Paola Contini Barbara Ruaro Alberto Sulli Stefano Scabini Emanuela Stratta Sabrina Paolino Carmen Pizzorni Vanessa Smith Renata Brizzolara
author_sort	Maurizio Cutolo
title	Effects of CTLA4-Ig treatment on circulating fibrocytes and skin fibroblasts from the same systemic sclerosis patients: an in vitro assay
title_short	Effects of CTLA4-Ig treatment on circulating fibrocytes and skin fibroblasts from the same systemic sclerosis patients: an in vitro assay
title_full	Effects of CTLA4-Ig treatment on circulating fibrocytes and skin fibroblasts from the same systemic sclerosis patients: an in vitro assay
title_fullStr	Effects of CTLA4-Ig treatment on circulating fibrocytes and skin fibroblasts from the same systemic sclerosis patients: an in vitro assay
title_full_unstemmed	Effects of CTLA4-Ig treatment on circulating fibrocytes and skin fibroblasts from the same systemic sclerosis patients: an in vitro assay
title_sort	effects of ctla4-ig treatment on circulating fibrocytes and skin fibroblasts from the same systemic sclerosis patients: an in vitro assay
publisher	BioMed Central
series	Arthritis Research & Therapy
issn	1478-6362
publishDate	2018-07-01
description	Abstract Background Systemic sclerosis (SSc) is characterized by vasculopathy and progressive fibrosis. CTLA4-Ig (abatacept) is able to interact with the cell surface costimulatory molecule CD86 and downregulate the target cell. The aim of this study was to evaluate the in-vitro effects of CTLA4-Ig treatment on circulating fibrocytes and skin fibroblasts isolated from the same SSc patient. Methods Circulating fibrocytes and skin fibroblasts were obtained from eight SSc patients with “limited” cutaneous involvement and from four healthy subjects (HSs). Samples were analyzed by fluorescence-activated cell sorter analysis (FACS) at baseline (T0) and after 8 days of culture (T8) for CD45, collagen type I (COL I), CXCR4, CD14, CD86, and HLA-DRII expression. Circulating fibrocytes were treated for 3 h and skin fibroblasts for 24/48 h with CTLA4-Ig (10, 50, 100, 500 μg/ml). Quantitative real-time polymerase chain reaction (qRT-PCR) was performed for CD86, COL I, FN, TGFβ, αSMA, S100A4, CXCR2, CXCR4, CD11a, and Western blotting was performed for COL I and FN. Results Using qRT-PCR, the T8-cultured SSc circulating fibrocytes which had not been treated with CTLA4-Ig showed higher gene expression for CD86, αSMA, S100A4, TGFβ, and COL I compared with HS circulating fibrocytes. Interestingly, αSMA/COL I gene expression was significantly lower only in the SSc circulating fibrocytes treated with CTLA4-Ig for 3 h (p < 0.01, p < 0.05). On the contrary, no effects were observed for either SSc or HS skin fibroblasts after CTLA4-Ig treatment. COL I and FN protein expression was unchanged in both SSc and HS skin fibroblasts by Western blot. Conclusions Circulating fibrocytes seem to be more responsive to CTLA4-Ig treatment than skin fibroblasts from the same SSc patient, likely due to their higher expression of CD86. CTLA4-Ig treatment might downregulate the fibrotic process in SSc patients by downregulating the fibrocytes, circulating progenitor cells.
topic	Fibrocytes Skin fibroblasts CTLA4-Ig Systemic sclerosis Connective tissue disease
url	http://link.springer.com/article/10.1186/s13075-018-1652-6
_version_	1612683788249202688

Effects of CTLA4-Ig treatment on circulating fibrocytes and skin fibroblasts from the same systemic sclerosis patients: an in vitro assay

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