| Budget Amount *help |
¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2001: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Research Abstract |
The presence of antigen-driven mechanism for production of auto antibodies has been suggested, based on their molecular characterization of CDR (complement determining region)-3 regions. In the present study, we tried to isolate auto-reactive B cells by flow cytometry, using B cell-specific monoclonal antibody, such as CD19, as well as auto-antigens including citrullination filaggrins and ribosomal P antigens. In the field of neurology, the well-characterized auto-antigens include acetyicholine receptors (AchR) in myasthenia gravis. However, AchR consists of several subunits, so we chose the rheumatoid arthritis autoantigen (citrullination filaggrins) and CNS-lupus autoantigens (ribosomal P antigens) for further experiments. The analysis of CDR3 regions of immunoglobulin genes from tonsil B cells were performed by RTPCR using total RNA from tonsils as template, specific primers (VH1-6) and isotype specific primer, which were subsequently cloned in TA vectors and sequenced by automated sequencer. However, we can not detect the autoreactive B cells by flow cytometry, even using the EAS method, which increase the sensitivity of flow cytometry. Our data suggests that the frequency of antireactive B cells in autoimmune disease might be very low.
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