Characterization of murine brain-reactive monoclonal IgG autoantibodies.

A diversity of brain-reactive autoantibodies (BRAA) is found in the sera and cerebrospinal fluid in elevated amounts in systemic lupus erythematosus (SLE) and correlates with some CNS manifestations of both human and murine SLE. In order to test the hypothesis that BRAA mediate the CNA manifestations, we are developing a library of brain-reactive monoclonal autoantibodies (BRMA) from autoimmune mice for characterization. We have recently reported on the production of BRMA of the IgM class from unimmunized autoimmune mice. In the current study we extend our investigations and report on the production and characterization of two BRMA of the IgG class. These antibodies react against integral brain membrane antigens of approximately 58, 32, and 30 kDa by immunoblot. Reactivity to such antigens is also found in a majority of the autoimmune murine serum samples tested, but not in nonautoimmune mice. These IgG BRMA show reactivity to cell bodies of the cerebral cortex, hippocampus, and hypothalamus of murine brain but not to fiber tracts. They also react with an integral thymus membrane antigen, but not to antigens of other tissues tested. Because of their properties, BRMA such as those characterized here are likely to be of pathogenic significance in CNS involvement in SLE.

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.