Antigen‐specific CD8+ T‐lymphocytes (cytotoxic T‐lymphocytes: CTL), as well as CD4+ T‐lymphocytes (helper T‐lymphocytes: Th), simultaneously play an important role in the elimination of intracellular bacteria such as Mycobacterium tuberculosis and Listeria monocytogenes. Administration of T‐cell epitope short peptide needs large numbers of peptides for effective vaccination due to its easily degradable nature in vivo. In this respect, biocompatible and biodegradable microparticles combined with CTL/Th‐hybrid epitope long peptide (long peptide) have been used to diminish the degradation of loaded peptide. The aim of this study is to develop a novel T cell‐oriented vaccine against intracellular bacteria that is composed of long peptide and poly (lactic‐co‐glycolic acid) (PLGA) microparticles. Mouse bone marrow‐derived dendritic cells (BMDCs) were loaded with L. monocytogenes listeriolysin O (LLO)‐derived or ovalbumin (OVA)‐derived long peptide/PLGA or other comparative antigens. The antigen‐loaded BMDCs were injected subcutaneously into the flank of mice twice, and then, the spleens were collected and lymphocyte proliferation and interferon‐γ production were evaluated. The median diameter of the PLGA spheres was 1.38 μm. Both LLO‐ and OVA‐long peptide/PLGA showed significantly more robust CTL and Th proliferations with higher interferon‐γ production than the long peptide alone or CTL and Th short peptides/PLGA vaccination. Furthermore, the LLO‐long peptide/PLGA vaccination showed a significantly lower bacterial burden in spleens compared with the long peptide alone or the CTL and Th short peptides/PLGA vaccination after the challenge of lethal amounts of L. monocytogenes. These results suggest that the novel vaccine taking advantages of CTL/Th‐hybrid epitope long peptide and PLGA microparticle is effective for protection against intracellular bacteria.