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. 2015 Jul 16;11(7):e1005041. doi: 10.1371/journal.ppat.1005041

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Fig 5 — A. PBMCs from 28 month old children (PROMOTE no chemoprevention control arm), malaria-exposed adults (PRISM cohort, Nagongera, Tororo District) and naïve adults were incubated for 7 days with protein extract from mature stage P. falciparum infected RBCs (PfSE) or uninfected RBCs (uE). T_reg frequencies were enumerated (FoxP3+CD25+CD127^dim) and induction factor was calculated based on frequency fold change between PfSE and uE stimulated PBMCs. Parasite induction of T_regs was reduced in exposed compared to naïve individuals. Wilcoxon signed rank test indicated for comparison between naïve-adults and exposed infant and adult samples. B. The proportion of children with any T_reg induction by PfSE was compared between those with low (<2 episodes ppy, n = 10) and high (>6 episodes ppy, n = 10) prior malaria incidence. There was a reduced proportion of infants with any T_reg induction in those who had high compared to low prior malaria incidence. Chi-square test indicated. C. Pro-apoptotic marker CD95 was measured on T_regs in children (PRISM cohort) from the high transmission area (Tororo District). Association between the level of CD95 expression on T_regs, as measured by MFI of CD95+ T_regs, and age was analyzed. The level of CD95 expression increased with age. D. Pro-survival Bcl2 was measured on T_regs in children (PRISM cohort, Nagongera, Tororo) from the high transmission area. Association between the expression of Bcl2 on the T_reg population and age was analyzed. The level of Bcl2 expression decreased with age. Spearman’s rho and p indicated. E. YoPro staining of T_regs from 28 month old children (PROMOTE, no chemoprevention control arm, with low (<2 episodes ppy) and high (>6 episodes ppy) prior malaria incidence was measured ex vivo and following stimulation with camptothecin (an activator of apoptosis) or P. falciparum antigen. Data from YoPro staining is representative of all measures of apoptosis (see S5 Fig additional measures of apoptosis including activated Caspase 3 and AnnexinV). There was no difference in sensitivity to apoptosis between infants with low and high prior malaria incidence. Wilcoxon signed rank test indicated.