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Plague is one of the most dangerous infections in humans caused by  Yersinia pestis , a Gram-negative bacterium. Despite of an overwhelming research success, no ideal vaccine against plague is available yet. It is well established that F1/LcrV based vaccine requires a strong cellular immune response for complete protection against plague. In our earlier study, we demonstrated that HSP70(II) of  Mycobacterium tuberculosis  modulates the humoral and cellular immunity of F1/LcrV vaccine candidates individually as well as in combinations in a mouse model. Here, we made two recombinant constructs  caf1–lcrV  and  caf1–lcrV–hsp70(II) . The  caf1  and  lcrV  genes of  Y. pestis  and  hsp70  domain II of  M. tuberculosis  were amplified by polymerase chain reaction. Both the recombinant constructs  caf1–lcrV  and  caf1–lcrV–hsp70(II)  were cloned in pET28a vector and expressed in  Escherichia coli.  The recombinant fusion proteins F1–LcrV and F1–LcrV–HSP70(II) were purified using Ni-NTA columns and formulated with alum to evaluate the humoral and cell mediated immune responses in mice. The protective efficacies of F1–LcrV and F1–LcrV–HSP70(II) were determined following challenge of immunized mice with 100 LD 50  of  Y. pestis  through intraperitoneal route. Significant differences were noticed in the titers of IgG and it’s isotypes, i.e., IgG1, IgG2b, and IgG3 in anti- F1–LcrV–HSP70(II) sera in comparison to anti-F1–LcrV sera. Similarly, significant differences were also noticed in the expression levels of IL-2, IFN-γ and TNF-α in splenocytes of F1–LcrV–HSP(II) immunized mice in comparison to F1–LcrV. Both F1–LcrV and F1–LcrV–HSP70(II) provided 100% protection. Our research findings suggest that F1–LcrV fused with HSP70 domain II of  M. tuberculosis  significantly enhanced the humoral and cellular immune responses in mouse model.

frontiers in microbiology

A Recombinant Trivalent Fusion Protein F1–LcrV–HSP70(II) Augments Humoral and Cellular Immune Responses and Imparts Full Protection against Yersinia pestis