Positive selection of the peripheral B cell repertoire in gut-associated lymphoid tissues

Abstract

Gut-associated lymphoid tissues (GALTs) interact with intestinal microflora to drive GALT development and diversify the primary antibody repertoire; however, the molecular mechanisms that link these events remain elusive. Alicia rabbits provide an excellent model to investigate the relationship between GALT, intestinal microflora, and modulation of the antibody repertoire. Most B cells in neonatal Alicia rabbits express V_Hn allotype immunoglobulin (Ig)M. Within weeks, the number of V_Hn B cells decreases, whereas V_Ha allotype B cells increase in number and become predominant. We hypothesized that the repertoire shift from V_Hn to V_Ha B cells results from interactions between GALT and intestinal microflora. To test this hypothesis, we surgically removed organized GALT from newborn Alicia pups and ligated the appendix to sequester it from intestinal microflora. Flow cytometry and nucleotide sequence analyses revealed that the V_Hn to V_Ha repertoire shift did not occur, demonstrating the requirement for interactions between GALT and intestinal microflora in the selective expansion of V_Ha B cells. By comparing amino acid sequences of V_Hn and V_Ha Ig, we identified a putative V_H ligand binding site for a bacterial or endogenous B cell superantigen. We propose that interaction of such a superantigen with V_Ha B cells results in their selective expansion.