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Role of intestinal microflora in diversification of the primary antibody repertoire in rabbit


, : Role of intestinal microflora in diversification of the primary antibody repertoire in rabbit. FASEB Journal 15(5): A1023, March 8

Germ-free rabbit studies and cecal ligation studies have demonstrated that GALT development and somatic diversification of VH genes in rabbits are dependent on interaction with intestinal flora. We examined antibody diversification in sterilely derived rabbits that were hand-raised in a setting that, although not germfree, precluded contact with conventional rabbits and their microflora. We found that 71% of the VDJ-Cmu genes cloned from these rabbits were undiversified, indicating that not simply intestinal contents, but specific members of the intestinal microflora are required for diversification of the rabbit primary antibody repertoire. To identify specific members of the intestinal microflora that can induce GALT development and somatic diversification, we examined appendix development in rabbits in which the appendix was ligated immediately after birth to prevent the entry of cecal contents. We introduced suspensions of cecal contents, ethanol-treated cecal contents, or various bacterial species singly or in various combinations into ligated appendices at 4-6 weeks of age and examined appendix development and VDJ gene diversification within the appendix two weeks later. Formation of proliferating B cell follicles was induced by total cecal contents, ethanol-treated cecal contents, and by a mixture of 6 bacteria. Bacteroides spp. alone induced B cell follicle formation without extensive proliferation and E. coli induced no follicle formation. Analysis of VDJ-Cmu genes from the ligated appendices demonstrated a correlation between the degree of somatic diversification and the extent of B cell proliferation within follicles. These results suggest that a certain subset of microbes is required for follicle formation and VDJ gene diversification in the rabbit appendix.

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