TY - JOUR
T1 - Induction of Inflammation and Fibrosis by Semaphorin 4A in Systemic Sclerosis
AU - Carvalheiro, Tiago
AU - Affandi, Alsya J
AU - Malvar-Fernández, Beatriz
AU - Dullemond, Ilse
AU - Cossu, Marta
AU - Ottria, Andrea
AU - Mertens, Jorre S
AU - Giovannone, Barbara
AU - Bonte-Mineur, Femke
AU - Kok, Marc R
AU - Marut, Wioleta
AU - Reedquist, Kris A
AU - Radstake, Timothy R
AU - García, Samuel
N1 - © 2019 The Authors. Arthritis & Rheumatology published by Wiley Periodicals, Inc. on behalf of American College of Rheumatology.
PY - 2019/10
Y1 - 2019/10
N2 - OBJECTIVE: To analyze the potential role of semaphorin 4A (Sema4A) in inflammatory and fibrotic processes involved in the pathology of systemic sclerosis (SSc).METHODS: Sema4A levels in the plasma of healthy controls (n = 11) and SSc patients (n = 20) were determined by enzyme-linked immunosorbent assay (ELISA). The expression of Sema4A and its receptors in monocytes and CD4+ T cells from healthy controls and SSc patients (n = 6-7 per group) was determined by ELISA and flow cytometry. Th17 cytokine production by CD4+ T cells (n = 5-7) was analyzed by ELISA and flow cytometry. The production of inflammatory mediators and extracellular matrix (ECM) components by dermal fibroblast cells (n = 6) was analyzed by quantitative polymerase chain reaction, ELISA, Western blotting, confocal microscopy, and ECM deposition assay.RESULTS: Plasma levels of Sema4A, and Sema4A expression by circulating monocytes and CD4+ T cells, were significantly higher in SSc patients than in healthy controls (P < 0.05). Inflammatory mediators significantly up-regulated the secretion of Sema4A by monocytes and CD4+ T cells from SSc patients (P < 0.05 versus unstimulated SSc cells). Functional assays showed that Sema4A significantly enhanced the expression of Th17 cytokines induced by CD3/CD28 in total CD4+ T cells as well in different CD4+ T cell subsets (P < 0.05 versus unstimulated SSc cells). Finally, Sema4A induced a profibrotic phenotype in dermal fibroblasts from both healthy controls and SSc patients, which was abrogated by blocking or silencing the expression of Sema4A receptors.CONCLUSION: Our findings indicate that Sema4A plays direct and dual roles in promoting inflammation and fibrosis, 2 main features of SSc, suggesting that Sema4A might be a novel therapeutic target in SSc.
AB - OBJECTIVE: To analyze the potential role of semaphorin 4A (Sema4A) in inflammatory and fibrotic processes involved in the pathology of systemic sclerosis (SSc).METHODS: Sema4A levels in the plasma of healthy controls (n = 11) and SSc patients (n = 20) were determined by enzyme-linked immunosorbent assay (ELISA). The expression of Sema4A and its receptors in monocytes and CD4+ T cells from healthy controls and SSc patients (n = 6-7 per group) was determined by ELISA and flow cytometry. Th17 cytokine production by CD4+ T cells (n = 5-7) was analyzed by ELISA and flow cytometry. The production of inflammatory mediators and extracellular matrix (ECM) components by dermal fibroblast cells (n = 6) was analyzed by quantitative polymerase chain reaction, ELISA, Western blotting, confocal microscopy, and ECM deposition assay.RESULTS: Plasma levels of Sema4A, and Sema4A expression by circulating monocytes and CD4+ T cells, were significantly higher in SSc patients than in healthy controls (P < 0.05). Inflammatory mediators significantly up-regulated the secretion of Sema4A by monocytes and CD4+ T cells from SSc patients (P < 0.05 versus unstimulated SSc cells). Functional assays showed that Sema4A significantly enhanced the expression of Th17 cytokines induced by CD3/CD28 in total CD4+ T cells as well in different CD4+ T cell subsets (P < 0.05 versus unstimulated SSc cells). Finally, Sema4A induced a profibrotic phenotype in dermal fibroblasts from both healthy controls and SSc patients, which was abrogated by blocking or silencing the expression of Sema4A receptors.CONCLUSION: Our findings indicate that Sema4A plays direct and dual roles in promoting inflammation and fibrosis, 2 main features of SSc, suggesting that Sema4A might be a novel therapeutic target in SSc.
KW - Adult
KW - Blotting, Western
KW - CD4-Positive T-Lymphocytes/immunology
KW - Case-Control Studies
KW - Cytokines/immunology
KW - Enzyme-Linked Immunosorbent Assay
KW - Extracellular Matrix/metabolism
KW - Female
KW - Fibroblasts/immunology
KW - Fibrosis/metabolism
KW - Humans
KW - Inflammation/immunology
KW - Male
KW - Microscopy, Confocal
KW - Middle Aged
KW - Monocytes/immunology
KW - Polymerase Chain Reaction
KW - Reverse Transcriptase Polymerase Chain Reaction
KW - Scleroderma, Systemic/immunology
KW - Semaphorins/metabolism
KW - Skin/cytology
KW - Th17 Cells/immunology
UR - http://www.scopus.com/inward/record.url?scp=85071254551&partnerID=8YFLogxK
U2 - 10.1002/art.40915
DO - 10.1002/art.40915
M3 - Article
C2 - 31012544
SN - 2326-5191
VL - 71
SP - 1711
EP - 1722
JO - Arthritis & rheumatology (Hoboken, N.J.)
JF - Arthritis & rheumatology (Hoboken, N.J.)
IS - 10
ER -