Detection of brain-reactive autoantibodies in the sera of autoimmune mice using ELISA

There are considerable problems with developing an assay to detect the often small quantities of autoantibodies which react against antigens in a heterogeneous and complex mixture from a source such as brain. An indirect enzyme-linked immunosorbent assay (ELISA) has been developed which can detect naturally occurring autoantibodies in serum that are reactive with integral brain membrane antigens. Sera were collected from autoimmune BXSB and NZB mice and non-autoimmune C57BL/6 mice at various ages and were assayed for the presence of brain-reactive autoantibodies (BRAAs). It is shown that this technique provides a highly sensitive, specific, and rapid assay for detecting BRAAs in serum. It shows that integral membrane antigens from whole brain can be isolated and used to detect and quantitate antibodies in the sera of autoimmune and non-autoimmune mice. The data also confirm studies, using different techniques, showing higher levels of autoantibodies to brain in autoimmune as compared to non-autoimmune mice. There are numerous potential applications for this ELISA, such as in rapidly screening large numbers of samples of biological fluids, tracking autoimmune disease progression over time, detecting small quantities of antibody against brain antigens, and as an assay system for investigating the role of BRAAs in the pathogenesis of immune mediated CNS disease.