Evaluation of CD4+/CD8 + T-cell expression and IFN-γ, perforin secretion for B–T constructs of F1 and V antigens of Yersinia pestis

SummaryYersinia pestis is a facultative bacterium that can survive and proliferate inside host macrophages and cause bubonic, pneumonic and systemic infection. Understanding the immune response generated by epitopes recognized by CD4 + and CD8 + T cells is important for the development of safe and effective vaccines designed to promote protective cellular immunity. Apart from humoral response, CD4 + T cells have shown to have a major role in combating the pneumonic form of the disease. In the present study, the secretion of IFN-γ and IL-4 by splenocytes, stimulated by different constructs of B and T cell epitopes of F1 and V antigens, was measured by ELISpot assay. We also measured perforin and IFN-γ expression as a function of cell mediated immunity by flow cytometry. Three B–T constructs of F1 and seven B–T constructs of V antigens produced a high number of IFN-γ secreting cells as compared to native antigen and a low number of IL-4 secreting cells. B–T conjugates of F1 and V antigens showed significantly high (p < 0.001) percentage of CD4 + IFN-γ+ cells as compared to CD8 + IFN-γ+ cells. Thus, the study highlights the importance of Th1 cytokine and existence of high proportion of CD4 + T cells probably contributing protection in the host. This study proposes a new perspective for the development of vaccination strategies for Y. pestis that trigger T cell immune response.

► F1 and V antigens are putative plague vaccine candidates.
► Subunit/peptide based vaccines can be made after identifying protective B and T cell epitopes of protein.
► Different B–T constructs of F1 and V antigens were synthesized.
► Cellular responses at systemic level and cytokine profile were studied in murine model using internasal immunization.
► FACS analysis proved expansion of CD4 + T cells and CD8 + T cells with IFN-γ, perforin and granzine were studied.