This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Antibiotic treatment-resistant Lyme arthritis is characterized by B-T cell aggregates and plasma cell clusters in synovial tissue from affected joints, after apparent eradication of Borrelia burgdorferi infection. It is suspected that this cellular activity in the joint is a due to activation of self-reactive B cells with subsequent production of autoantibodies. This work focuses on identification of potential self antigenic proteins from synovial tissue. Microdissected plasma cells from affected joints were cultured in vitro and the antibodies were purified from the cell supernates. These potential autoantibodies were conjugated to agarose beads and used to affinity purify antigenic proteins from synovial fluid. Proteins were eluted from the beads and separated by SDS-PAGE. Protein bands were excised from gels and subjected to in-gel trypsin digestion. Peptides extracted from the in-gel digests were analyzed by MALDI-TOF mass spectrometry for protein identification. We have identified a cytokeratin that was present in one sample, but not in control blank gel slices. We are currently in the process of confirming the finding by western blot as well as continuing to search for other proteins in additional samples.
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