Tight-skin mouse autoantibody repertoire: analysis of VH and VK gene usage

Abstract

In the previous studies we have shown that tight-skin (TSK) mouse is an experimental model for systemic sclerosis. This mutant mouse develops autoantibodies specific for scleroderma target antigens. To determine whether the expansion of autoantibody repertoire in TSK mouse occurs by selective expansion of certain variable region gene families, and whether CD5+ B cells contribute significantly to the production of autoantibodies, we have analyzed a panel of 60 hybridomas producing autoantibodies specific for scleroderma target autoantigens. Northern analysis of RNAs from these hybridomas showed that 70% were expressing genes from VHJ558 family while genes from 36-09 and J606 families were not at all represented. In contrast, in the cDNA libraries derived from splenic B cells, the expression of VHJ558 and 36-09 gene families were at an expected frequency corresponding to their genomic complexity (44% and 11.6%, respectively). These results demonstrate that there is a strong bias toward the use of J558 genes in TSK mouse autoantibody repertoire. On the other hand the expression of VK gene families was mostly random and corresponded to their frequency in splenic C kappa cDNA library. The pairing of VH:VK genes was stochastic. Analysis of the expression of J segments, however, revealed that JH2 and JK2 were predominantly used in the autoantibodies. Analysis of the expression CD5 mRNA in these hybridomas indicate that CD5+ B cells do not contribute significantly to the autoimmunity in TSK mice. These findings suggest that the expansion of peripheral autoreactive B cells in TSK mouse is determined by their immunoglobulin variable region rather than the genetic properties linked to a particular B cell subset.