A study of antigen selection by extracellular vesicles as vaccine candidates against Mycobacterium tuberculosis infection

Introduction. Tuberculosis (TB), an infectious disease caused by Mycobacterium tuberculosis ( M. tb ), remains a significant global public health concern. It is crucial to develop more effective vaccines for TB in order to achieve global control of the disease. Extracellular vesicles (EVs) are spherical membrane-bound structures released by pathogens and host cells. During the course of an infection, both pathogen- and host-derived EVs are produced and play important roles in determining the course of the infection. EVs offer intriguing tools as potential vaccines due to their ability to deliver multiple pathogen or host antigens. Hypothesis /Gap Statement. We hypothesized that EVs derived from M. tb and EVs from M. tb -infected macrophages may serve as potential vaccine candidates against M. tb infection. Aim . This study aims to compare the immunogenicity and immune protection between M. tb EVs and M. tb -infected macrophage-derived EVs. Methodology. In this study, EVs were extracted from culture supernatants of M. tb and M. tb -infected macrophages, respectively. Mass spectrometry was employed to explore the antigen composition of H37Rv-Mφ-EVs and H37Rv-EVs. Cytokine profiling and antibody response assays were used to analyse the immunogenicity offered by EVs. Additionally, we used histological examination to evaluate and protective efficacy of the EVs. Results. Our results demonstrated that mice immunized by EVs released from M. tb -infected macrophages induced stronger inflammatory cytokine response than M. tb . Moreover, EVs from M. tb -infected macrophages reinforced T-cell activation and antibody response compared to M. tb EVs. Proteomic analysis revealed that EVs from M. tb -infected macrophages containing immunodominant cargos have stronger binding ability with major histocompatibility complex molecules, which may contribute to the protection from M. tb infection. Indeed, immunization of EVs released from M. tb -infected macrophages significantly reduced the bacterial load and better protection against M. tb infection than EVs from M. tb . Importantly, the selected antigens (Ag85B, ESAT-6 and the Rv0580c) from EVs of M. tb -infected macrophages exhibited effective immunogenicity. Conclusion. Our results suggested that EVs derived from M. tb -infected macrophages might serve as a proper antigenic library for vaccine candidates against M. tb challenge.