Chimeric antigen carried by extracellular vesicles induces stronger protective immunity against Mycobacterium tuberculosis infection

Although Bacillus Calmette-Guerin (BCG) has been used in human for centuries, tuberculosis (TB) remains one of the deadliest infectious diseases. There have been remarkable successes in the field of TB vaccine research over the past decade, but the search for a better vaccine candidate is still a challenge. Extracellular vesicles (EVs) possess a multitude of properties that make them attractive candidates for the development of novel, cell-free, non-replicative, and safe vaccine system. These properties include their small size, inherent immunogenicity, ability to be taken up by immune cells, self-adjuvant capability and the comprehensive distribution of concentrated antigens. In this study, we designed a newly chimeric antigen TB vaccine (CA) with three Mycobacterium tuberculosis (M. tb) antigens that identified from extracellular vesicle derived from M. tb-infected macrophage. We confirmed that the CA stimulated a more pronounced immune response and enhanced T-cell activation, thereby providing superior protection against Mycobacterium tuberculosis infection in comparison to the bivalent antigens. Importantly, the EVs carrying CA (EVs-CA) provided enhanced protection against M. tb infection compared to unencapsulated CA antigen. Moreover, we established an EV-carried CA system (EVs-CA) and released from a transformed cell line using endogenous loading of antigen method. This method displayed the CA could efficiently package into EVs and increased concentration of this antigen. The chimeric antigen carried by EVs induced higher levels of cytokines production and specific cytotoxic T lymphocytes, resulted in enhancing antibody response and improving protective efficacy. Our findings suggested that the potential of EVs as delivery system to carry the M. tb-specific chimeric antigen for controlling Mycobacterium tuberculosis infection.