Figure S1.

Systemic inflammation is not attenuated upon rechallenge. (A) Data were extracted from Gonçalves et al. (2014) to examine the frequency of severe or complicated malaria during the first 14 infections of life in infants living in a hyperendemic setting. We first plotted the total number of cases of malaria (including mild or uncomplicated episodes) and used least squares regression to impute missing values (for infection numbers 9, 11, 12, and 13). Imputed values are shown as a grey triangle whereas filled circles indicate data as reported by Gonçalves et al. The total number of children experiencing at least one episode of malaria was n = 715. (B) We next plotted the incidence of severe or complicated malaria at each order of infection (as shown in Fig. 1, A and B) and performed maximum likelihood estimation to select the best model fit for these data; log likelihood (logL) and AIC both show that an exponential decay in risk provides the best fit (SE, standard error). (C) Healthy malaria-naive adults were enrolled in the VAC063 study and infected up to three times with P. falciparum (clone 3D7) by direct blood challenge. The mean frequency of eosinophils, lymphocytes, monocytes, and neutrophils in whole blood at baseline and diagnosis is shown (note that the loss of lymphocytes at diagnosis is comparable in all three infections). (D) RNAseq was used to identify differentially expressed genes in whole blood at diagnosis (versus baseline) (adj P < 0.05 and >1.5 fold-change). Volcano plots show all differentially expressed genes (colored dots) and the dashed lines represent the significance/fold-change (FC) cutoffs. The top 10 differentially expressed genes (lowest adj P) in each infection are labeled (first, second, and third infection were analyzed independently). (E) The log₂ fold-change of signature genes associated with interferon signaling and type I inflammation are shown at diagnosis (versus baseline) in the first, second, and third infection. Square brackets indicate that common protein names have been used. In C, n = 10 (first and second infection) and n = 6 (third infection). In D and E, n = 10 (first infection), n = 9 (second infection), and n = 6 (third infection). v1040 was excluded from RNAseq analysis in the second infection because their baseline sample failed QC.