Overexpression and secretion of the soluble CTLA-4 splice variant in various autoimmune diseases and in cases with overlapping autoimmunity

Abstract

Aim: To explore the potential genetic association of CTLA-4 Exon1 +49A/G and 3'UTR (AT)(n) to susceptibility to systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and overlapping (OP) autoimmunity; affected with more than one autoimmune disease. Expression of two major CTLA-4 isoforms; full length (mCTLA-4) and soluble (sCTLA-4) were explored in all subjects. A total of 680-age/gender/ethnically matched Kuwaitis were recruited and polymerase chain reaction (PCR)-fragment analysis was employed for genotyping both markers. mCTLA-4 and sCTLA-4 mRNA expression were analyzed using quantitative real time-PCR. The enzyme-linked immunosorbent assay (ELISA) was used to screen sCTLA-4 in all subjects' sera.

Results: Only two CTLA-4 3'UTR (AT)(n) allelotypes; (AT)(15) and (AT)(6) were detected. The heterozygous (AT)(15/6) genotype confers protectivity rather than susceptibility to SLE (p=0.01, odds ratio=0.43, and confidence interval=0.21-0.86). No significant association was observed between Exon 1 +49A/G and any of the tested diseases. A consistently high serum sCTLA-4 level was observed in RA (6.8 ng/mL, p=0.005), SLE (6.34 ng/mL, p=0.007), and OP (8.75 ng/mL, p=0.012) compared to healthy control. A significant increase in the expression of sCTLA-4 mRNA was observed in OP (p=0.05) and SLE (p=0.047), while a significant increase in the expression of mCTLA-4 (p=0.01) was observed only in OP.

Conclusion: The present study is the first to report a statistically significant association between OP and serum sCTLA-4. The novelty of our study is the significance of CTLA-4 in the pathogenesis of OP besides SLE and RA.

FIG. 1.

Serum soluble form CTLA-4 (sCTLA-4) found in autoimmune patients, such as rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), overlapping (OP), and healthy control (HC). The concentration of serum sCTLA-4 in a total of 134 autoimmune patients and 68 healthy controls were analyzed using the ELISA test. Results are expressed in ng/mL.