Dose of Incorporated Immunodominant Antigen in Recombinant BCG Impacts Modestly on Th1 Immune Response and Protective Efficiency againstMycobacterium tuberculosisin Mice

Abstract

One approach for improving BCG efficacy is to utilize BCG as vehicle to develop recombinant BCG (rBCG) strains overexpressingMycobacterium tuberculosis (M. tb)antigens. Also expression level of a candidate antigen should impact the final T cell responses conferred by rBCG. In this study, based on our previously constructed differential expression system, we developed two rBCG strains overexpressingM. tbchimeric antigen Ag856A2 (coding a recombinantag85awith 2 copies ofesat-6inserted atAccI site ofag85a) at differential levels under the control of the subtly modifiedfurApromoters. These two rBCG strains were used to vaccinate C57BL/6 mice and exploit dose of incorporated antigen in rBCG to optimize immune response and protective efficiency againstM. tbchallenge in mouse model. The results showed that rBCG strains overexpressing Ag856A2 at differential levels induced different antigen-specific IFN-γproduction and comparable number ofM. tb-specific CD4 T cells expressing IL-2.M. tbchallenge experiment showed that rBCG strains afforded enhanced but comparable immune protection characterized by reduced bacillary load, lung pathology, and inflammation. These results suggested that the dose of antigens incorporated in rBCG can impact T cell immune responses but imposed no significantly differential protective efficacies.