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Comparative Analysis of the CD19⁺ and CD138⁺ Cell Antibody Repertoires in the Cerebrospinal Fluid of Patients with Multiple Sclerosis¹

Alanna M. Ritchie^*, Donald H. Gilden^*,, R. Anthony Williamson, Mark P. Burgoon^*, Xiaoli Yu^*, Karen Helm, John R. Corboy^* and Gregory P. Owens^2,*

Departments of ^* Neurology and Microbiology and University of Colorado Cancer Center, University of Colorado Health Sciences Center, Denver, CO 80262; and Department of Immunology, The Scripps Research Institute, La Jolla, CA 92037

Increased amounts of intrathecally synthesized IgG and oligoclonal bands have long been recognized as a hallmark of multiple sclerosis (MS). B cells and plasma cells are components of the inflammatory infiltrates in both active and chronic MS lesions, and increased numbers of these cells are present in MS cerebrospinal fluid (CSF). Single-cell RT-PCR was used to analyze both the CD19⁺ B cell and CD138⁺ plasma cell populations in CSF of two patients with clinically definite MS and of one MS patient whose CSF was obtained after a clinically isolated syndrome, but before the second episode. Sequence analysis of amplified IgG V region sequences identified the rearranged germline segments, extent of somatic mutation, and clonal relationships within and between the two cell populations in the three MS patients. Expanded B cell and plasma cell clones were detected in each MS CSF and in all three patients the CD138⁺ IgG repertoire was more restricted. However, little if any significant sequence overlap was observed between the CD19⁺ and CD138⁺ repertoires of each donor. Detection of plasma cell clones by single-cell PCR will facilitate the in vitro production of recombinant Abs useful in identifying disease-relevant Ags.

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