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Cuong Nguyen, Adebola Ogunniyi, Christopher Love, Characterization of B cell repertoires in Sjögren’s syndrome by microengraving (44.48), The Journal of Immunology, Volume 186, Issue 1_Supplement, April 2011, Page 44.48, https://doi.org/10.4049/jimmunol.186.Supp.44.48
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Abstract
Microengraving is a novel technology that uses an array of microfabricated wells to isolate and characterize large numbers of single cells. Using printing technique, secreted antibodies can be captured and characterized on glass slides. Using microengraving, we profiled the antigenic repertoires of B cells reacting against salivary gland (SG) tissues in Sjögren’s syndrome (SjS), an autoimmune disease targeting the exocrine glands. Cells from spleens and cervical lymph nodes (CLN) of normal C57BL/6 (B6) and SjS-susceptible (SjSs) C57BL/6.NOD-AecAec2 mice were isolated and loaded onto microwells. Capture slides preincubated with mouse Igs were used for printing. Detection antibodies included IgG1, IgG3, and SG lysates of B6 and SjSs mice that were labeled with different fluorescent dyes. Data indicated an increase in the frequency of IgG1 antibodies against SG lysates of B6 and SjSs mice when printed using CLN cells of SjSs mice but not B6 mice. In contrast, an increase in the frequency of IgG3 antibodies against SG tissues of B6 and SjSs mice was only observed when printed with SjSs mice splenocytes. These data demonstrate the diversity in the isotypic repertoire of antibodies during autoimmune process, and further revealed a dichotomy in peripheral tolerance. The result supports the generation of self-reactive B cell repertories during the autoimmune process. Furthermore, the use of micrograving of single cells might allow for identification of novel biomarkers.
