| Full text | |
| Author(s): |
Keppeke, Gerson Dierley
;
Satoh, Minoru
;
Kayser, Cristiane
;
Matos, Pedro
;
Hasegawa, Tomoko
;
Tanaka, Shin
;
Diogenes, Larissa
;
Amaral, Rogerio Quintiliano
;
Rodrigues, Silvia Helena
;
Andrade, Luis Eduardo Coelho
Total Authors: 10
|
| Document type: | Journal article |
| Source: | FRONTIERS IN IMMUNOLOGY; v. 13, p. 11-pg., 2022-09-26. |
| Abstract | |
Anti-fibrillarin autoantibodies are useful for the diagnosis and prognosis of systemic sclerosis (SSc). Anti-fibrillarin produces a clumpy nucleolar pattern in indirect immunofluorescence assay on HEp-2 cells (HEp-2 IFA). Here we develop and validate a reliable cell-based anti-fibrillarin assay (Fibrillarin/CBA) for use in clinical diagnostic laboratories. A TransMembrane Signal was fused to the human fibrillarin gene (TMS-fibrillarin). HEp-2 cells overexpressing transgenic TMS-fibrillarin at the cytoplasmic membrane were used as IFA substrate in the Fibrillarin/CBA. Sixty-two serum samples with nucleolar pattern in the HEp-2 IFA (41 clumpy; 21 homogeneous/punctate) were tested for anti-fibrillarin using Fibrillarin/CBA, immunoprecipitation (IP), line-blot and ELISA. In addition, samples from 106 SSc-patients were evaluated with Fibrillarin/CBA and the results were correlated with disease phenotypes. Thirty-eight of 41 samples with the clumpy nucleolar pattern (92.7%) were positive in the Fibrillarin/CBA, while all 21 samples with other nucleolar patterns were negative. Fibrillarin/CBA results agreed 100% with IP results. Among the 38 Fibrillarin/CBA-positive samples, only 15 (39.5%) and 11 (29%) were positive for anti-fibrillarin in line-blot and ELISA, respectively. Higher frequency of diffuse cutaneous SSc (dcSSc) phenotype (72.7% vs 36.8%; p=0.022), cardiac involvement (36.4% vs 6.5%; p=0.001) and scleroderma renal crisis (18.2% vs 3.3% p = 0.028) was observed in SSc patients with positive compared to negative Fibrillarin/CBA result. Performance of Fibrillarin/CBA in the detection of anti-fibrillarin autoantibodies was comparable to the gold standard IP. Positive Fibrillarin/CBA results correlated with disease phenotypes known to be associated with anti-fibrillarin autoantibodies, underscoring the clinical validation of this novel assay. (AU) | |
| FAPESP's process: | 17/20745-1 - Autoantibodies depletion on autoreactive plasmocytes by genome editing with the CRISPR/Cas9 system |
| Grantee: | Gerson Dierley Keppeke |
| Support Opportunities: | Research Grants - Young Investigators Grants |
| FAPESP's process: | 21/04588-9 - A cell-based assay's construction for the detection of acetylcholine receptor |
| Grantee: | Larissa Diogenes Santos |
| Support Opportunities: | Scholarships in Brazil - Master |