PD-1 deficiency promotes TFH cells expansion in ITV-immunized mice by upregulating cytokines secretion

Background: T follicular helper (TFH) cells are fundamental for the development of humoral immunity. In our previous study, we found that PD-1 deficiency substantially promoted the expansion of Plasmodium-specific TFH cells and enhanced the humoral immunity of ITV (infection treatment vaccine)-immunized mice. However, the underlying mechanism by which PD-1 signaling modulates TFH cells activation remains unclear. Methods: Mice were immunized with the ITV following the standard procedures. The activation phenotype of CD11c(+)CXCR5(+) dendritic cells (DCs), the frequency and number of splenic follicular regulatory T cells (TFR cells), Plasmodium-specific TFH cells and germinal center (GC) B cells were analyzed by FACS. The levels of serum cytokines were quantified using the cytometric bead array (CBA) and in vivo cytokine neutralization was carried out according to a previously described protocol and verified by serum cytokine detection. Results: We found that PD-1(-/-) naive and immunized mice had more TFR cells in the spleen than WT and WT immunized mice. Additionally, CXCR5(+) DC, which prime TFH cells, were activated at similar levels in ITV-immunized WT and PD-1(-/-) mice. However, the serum levels of IL-10, IFN-gamma and MCP-1 were significantly increased in ITV-immunized PD-1(-/-) mice, and treatment with an anti-IL-10, anti-IFN-gamma or anti-MCP-1 neutralizing antibody in vivo markedly impaired the development of TFH cells and GC B cells. Conclusions: Our findings demonstrate that the modulation of TFH cells by PD-1 signaling is dependent on the cytokines IL-10, IFN-gamma and MCP-1 in ITV-immunized mice. These results could facilitate the design of an effective malaria vaccine with the aim of inducing humoral immune responses.