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The efficacy of most subunit vaccines is limited by the induction of weak T cell immunity. The use of multiple antigen delivery platforms represents a strategy to overcome such limitations. In this study, we co-expressed the major Bacillus Calmette-Guérin (BCG) vaccine antigens Ag85B and ESAT-6 in Mycobacterium smegmatis and constructed a BCG live vaccine candidate by co-expressing Ag85B and ESAT-6 with the MV-vectored Ag85B/ESAT-6 fusion protein. The results demonstrated that the fusion protein increased the expression of MV-vaccine components Ag85B and ESAT-6 in BCG, promoted the uptake of BCG in macrophages, and induced strong immune responses in
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