The flagellin D2/D3 domain induces TLR5- and inflammasome- independent antibody responses (INM2P.357)

Abstract

Abstract The detailed understanding of the molecular mechanisms underlying the biological activity adjuvants is critical for rational design of more effective vaccines. Bacterial flagellin, a potent immunogen, is being developed as a new adjuvant and platform for human vaccines. We have demonstrated that the conserved D0 and D1 domains of flagellin promote antibody responses through activation of their respective innate immune receptors, but are not required for Salmonella flagellin (FliC) induced antibody responses. We hypothesized that additional flagellin domains also influence specific aspects of flagellin-induced B cell responses. Mice were immunized with a mixture of FliC mutants proteins and ovalbumin, and sera were collected for quantification of anti-flagellin and anti-ovalbumin specific antibody responses. The results from our studies demonstrate that the D2/D3 domain of the FliC is also required for flagellin-induced primary antibody responses and robust secondary responses. D2/D3 domain is capable of inducing antibody responses independent of TLR5 and the inflammasome. Our data indicate the flagellin D2/D3 domain synergizes with the TLR5 (D1) and inflammasome (D0) activating domains to induce potent antibody responses, and that the D2/D3 domain also functions as an adjuvant to promote antibody responses against co-administered antigens. Future studies will elucidate the contribution of the FliC D2/D3 domain to B cell responses and its utility in vaccines.