An unadjuvanted model for oral immunization revealing complex regulatory control of gut IgA responses to soluble protein

Abstract

We have identified a unique mouse model in which oral immunization with soluble protein, without a potent mucosal adjuvant, is highly effective at stimulating strong gut IgA responses. In fact, cholera toxin (CT), which is the gold standard for oral adjuvants, did not enhance the gut IgA response to ovalbumin (OVA) in this model. As we dissected the mechanism allowing for an unadjuvanted strong gut IgA response we found that adoptively transferred OVA-peptide-specific TCR Tg DO11.10 CD4 T cells into nude Balb/c mice were sufficient to promote the gut IgA response. However, only CD4 T cells with a rearranged TCR a-chain, reactive to unknown, but possibly microbial, antigens in the gut, were effective, because SCID DO11.10 CD4 T cells failed to support a gut IgA response. Co-transfer of wild-type CD4 T cells with DO11.10 CD4 T cells into nude Balb/c hosts completely obliterated the response, but not if the CD4 T cells were isolated from IL-10−/− mice. Hence, the gut IgA response was under strict regulatory CD4 T cell control. By contrast, if CT adjuvant was admixed with OVA we completely rescued the gut IgA response following oral immunization. Thus, oral CT adjuvant primarily exerted a modulating effect on the regulatory CD4 T cell control in Peyer’s patches (PP), impacting on the balance between follicular helper and regulatory T cells. While both Th17 and CD25⁺ Tregs, have been claimed sufficient for supporting gut IgA responses, our model clearly suggests that the regulatory environment in PP is more complex, as will be discussed.