Fig. 5.

PMIF neutralization decreases inflammatory cytokine production and enhances the development of CD4 T cells into effector memory and memory precursors during blood-stage infection. a, b Serum levels of the indicated cytokines were detected by specific ELISA 7 days after injection of 10⁶ PbAWT iRBCs in mice immunized with RNA replicons encoding Con RNA or PMIF RNA. Data are representative of two independent experiments. Bars represent the mean of 6 mice ± SD; *p < 0.05,**p < 0.01 by Mann–Whitney test. c On day 7, 10, and 15 after infection, splenocytes were isolated and stimulated ex vivo with iRBC lysates in the presence of Brefeldin A. Representative dot plots and frequencies of PbAWT responsive CD4 T cells (Ki67⁺CD4⁺) expressing IFN-γ in spleens was detected by intracellular staining and analyzed by flow cytometry. Data are representative of two independent experiments. Bars represent the mean of 10 mice ± SD; *p < 0.05 by Mann–Whitney test. d, e Numbers of PbAWT responsive CD4 T cell (Ki67⁺CD4⁺) subsets, including T effector (Teff): CD62L⁻IL7Rα⁻, T effector memory (Tem): CD62L⁻IL7Rα⁺, and T memory (Tmem): CD62L⁺IL7Rα⁺ at day 7 and 15 after infection. The contribution of each memory CD4 T cell subset is expressed relative to the total number of PbAWT responsive CD4 T cells. Data are representative of two independent experiments. Bars represent the mean of 10 mice ± SD; n.s.: non-significant, *p < 0.05, **p < 0.001, ^#p < 0.0001 by Mann–Whitney test