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Department of Microbiology, College of Natural Sciences, Kangwon National University, Chunchon, Korea, and
Laboratory of Mucosal Immunology, Department of Medicine, University of California, San Diego, La Jolla, CA 92093
Cholera toxin (CT) and its B subunit (CTB) are potent immunogens and adjuvants that, either alone or linked to protein Ags, can stimulate mucosal immune responses, modulate the induction of oral tolerance, and stimulate IgA isotype switching. The present studies addressed the mechanisms by which CT and CTB promote IgA switching. CT and rCTB, in the presence of IL-2, significantly increased IgA isotype switching at the clonal level in populations of purified and LPS-activated murine surface IgA- spleen B cells, as determined by ELISA, enzyme linked immunospot assays, and limiting dilution analysis. The IgA stimulatory effects of CT and CTB were independent of the A subunit of CT. CTB and CT did not increase the secretory rate of IgA-producing cells or the clonal burst size of IgA clones, and did inhibit B cell growth. Because TGF-ß1 also inhibits B cell growth and promotes IgA switching, further studies tested whether the activity of CTB and CT on IgA isotype switching was mediated through TGF-ß1. Anti-TGF-ß Ab and soluble TGF-ß1 type IIR inhibited CTB- and CT-stimulated IgA isotype switching. Furthermore, increased TGF-ß1 mRNA levels and bioactive TGF-ß1, within a range shown to induce IgA isotype switching, were detected in cultures of surface IgA- B cells stimulated with CT or CTB and IL-2. These data indicate that CTB- and CT-stimulated IgA isotype switching are mediated through TGF-ß1. The finding that CTB up-regulates TGF-ß1 activity has important implications for understanding the mechanisms by which CTB promotes both IgA mucosal immunity and oral tolerance.
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