A multi-epitope vaccine incorporating adhesin-derived antigens protects againstinfection and dissemination.

INTRODUCTION: Adhesion to host cells is the first and essential step in() infection. Among adhesion molecules, the PGRS domain of PE_PGRS33 plays a critical role in invasion but is dominated by B cell epitopes and lacks sufficient T cell epitopes, restricting its capacity to induce a balanced immune response.

METHODS: To overcome this limitation, we employed an integrative reverse vaccinology pipeline combining computational prediction and experimental validation. Helper and cytotoxic T lymphocyte epitopes were incorporated from multipleadhesins as well as other virulence-associated proteins, and adjuvant sequences were systematically evaluated in silico.

RESULTS: Among three multi-epitope constructs, the Toll-like receptor 2 (TLR2)-agonist and pan HLA DR-binding epitope (PADRE)-adjuvanted vaccine (TLR2-vaccine) emerged as the most promising candidate. In murine models, TLR2-vaccine induced strong antigen-specific antibody and IFN-γ responses, significantly reduced bacterial loads following H37Ra challenge, and effectively prevented extrapulmonary dissemination.

DISCUSSION: These findings highlight the potential of adhesin-inclusive multi-epitope vaccines to elicit both humoral and cellular immunity and demonstrate how computational vaccinology can accelerate the development of targeted interventions against tuberculosis.