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Antibodies against specific extractable nuclear antigens (ENAs) as diagnostic and prognostic tools and inducers of a profibrotic phenotype in cultured human skin fibroblasts: are they functional?
- Source :
- Arthritis Research & Therapy, Vol 21, Iss 1, Pp 1-11 (2019), Arthritis Research & Therapy
- Publication Year :
- 2019
- Publisher :
- BMC, 2019.
-
Abstract
- Background The importance of systemic sclerosis (SSc) autoantibodies for diagnosis has become recognized by their incorporation into the 2013 ACR/EULAR classification criteria. Clear prognostic and phenotypic associations with cutaneous subtype and internal organ involvement have been also described. However, little is known about the potential of autoantibodies to exert a direct pathogenic role in SSc. The aim of the study is to assess the pathogenic capacity of anti-DNA-topoisomerase I (anti-Topo-I) and anti-centromeric protein B (anti-Cenp-B) autoantibodies to induce pro-fibrotic markers in dermal fibroblasts. Methods Dermal fibroblasts were isolated from unaffected and affected skin samples of (n = 10) limited cutaneous SSc (LcSSc) patients, from affected skin samples of diffuse cutaneous (DcSSc) patients (n = 10) and from healthy subjects (n = 20). Fibroblasts were stimulated with anti-Topo-I, anti-Cenp-B IgGs, and control IgGs in ratios 1:100 and 1:200 for 24 h. Cells were also incubated with 10% SSc anti-Topo-I+ and anti-Cenp-B+ whole serum and with 10% control serum for 24 h. Viability was assessed by MTT test, while apoptosis was assessed by flow cytometry. Activation of pro-fibrotic genes ACTA2, COL1A1, and TAGLN was evaluated by quantitative real-time PCR (qPCR), while the respective protein levels alpha-smooth-muscle actin (α-SMA), type-I-collagen (Col-I), and transgelin (SM22) were assessed by immunocytochemistry (ICC). Results MTT showed that anti-Cenp-B/anti-Topo-I IgGs and anti-Cenp-B+/anti-Topo-I+ sera reduced viability (in a dilution-dependent manner for IgGs) for all the fibroblast populations. Apoptosis is induced in unaffected LcSSc and control fibroblasts, while affected LcSSc/DcSSc fibroblasts showed apoptosis resistance. Basal mRNA (ACTA2, COL1A1, and TAGLN) and protein (α-SMA, Col-1, and SM22) levels were higher in affected LcSSc/DcSSc fibroblasts compared to LcSSc unaffected and to control ones. Stimulation with anti-Cenp-B/anti-Topo-I IgGs and with anti-Cenp-B+/anti-Topo-I+ sera showed a better induction in unaffected LcSSc and control fibroblasts. However, a statistically significant increase of all pro-fibrotic markers is reported also in affected LcSSc/DcSSc fibroblasts upon stimulation with both IgGs and sera. Conclusions This study suggests a pathogenic role of SSc-specific autoantibodies to directly induce pro-fibrotic activation in human dermal fibroblasts. Therefore, besides the diagnostic and prognostic use of those autoantibodies, these data might further justify the importance of immunosuppressive drugs in the early stages of the autoimmune disease, including SSc. Electronic supplementary material The online version of this article (10.1186/s13075-019-1931-x) contains supplementary material, which is available to authorized users.
- Subjects :
- 0301 basic medicine
Adult
Male
lcsh:Diseases of the musculoskeletal system
Extractable nuclear antigens
Cell Survival
Immunocytochemistry
Apoptosis
Flow cytometry
Centromeric protein B
03 medical and health sciences
0302 clinical medicine
Fibrosis
medicine
Humans
Fibroblast
Cells, Cultured
Aged
Skin
Autoantibodies
030203 arthritis & rheumatology
Autoimmune disease
Scleroderma, Systemic
medicine.diagnostic_test
biology
business.industry
Autoantibody
Antigens, Nuclear
Middle Aged
Fibroblasts
medicine.disease
Prognosis
Immunohistochemistry
Topoisomerase I
030104 developmental biology
medicine.anatomical_structure
Systemic sclerosis, Fibrosis, Autoantibodies, Fibroblasts, Centromeric protein B, Topoisomerase I
Immunology
biology.protein
Systemic sclerosis
Female
Antibody
lcsh:RC925-935
business
Research Article
Subjects
Details
- Language :
- English
- ISSN :
- 14786362
- Volume :
- 21
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Arthritis Research & Therapy
- Accession number :
- edsair.doi.dedup.....7b4683c3aca07074ca111d421d40ef4c