Abstract
Cerebral malaria (CM) is a severe complication of Plasmodium falciparum infection responsible for thousands of deaths in children in sub-Saharan Africa. CM pathogenesis remains incompletely understood but a number of effectors have been proposed, including plasma microparticles (MP). MP numbers are increased in CM patients’ circulation and, in the mouse model, they can be localised within inflamed vessels, suggesting their involvement in vascular damage. In the present work we define, for the first time, the protein cargo of MP during experimental cerebral malaria (ECM) with the overarching hypothesis that this characterisation could help understand CM pathogenesis. Using qualitative and quantitative high-throughput proteomics we compared MP proteins from non-infected and P. berghei ANKA-infected mice. More than 360 proteins were identified, 60 of which were differentially abundant, as determined by quantitative comparison using TMTTM isobaric labelling. Network analyses showed that ECM MP carry proteins implicated in molecular mechanisms relevant to CM pathogenesis, including endothelial activation. Among these proteins, the strict association of carbonic anhydrase I and S100A8 with ECM was verified by western blot on MP from DBA/1 and C57BL/6 mice. These results demonstrate that MP protein cargo represents a novel ECM pathogenic trait to consider in the understanding of CM pathogenesis.
Highlights
Cerebral malaria (CM) is a severe complication of Plasmodium falciparum infection responsible for thousands of deaths in children in sub-Saharan Africa
Host-derived plasma microparticles (MP) are well recognised as being involved in the pathogenesis of both human and experimental CM17,18,20,21, the contribution of their protein cargo to the pathological mechanisms leading to this syndrome has not been deciphered yet
This is the first time that this strategy is applied to experimental cerebral malaria (ECM) and that the protein cargo of MP obtained from individual mice is successfully investigated by Tandem Mass TagTM (TMT) quantitative proteomics
Summary
In the TMT0 experiment we identified 184 NI MP proteins and 164 ECM MP proteins (2 unique peptides, FDR ≤1%, Fig. 3A). Considering the two TMT0 and the two TMT6 experiments together, we globally identified 368 murine plasma MP proteins (2 unique peptides, FDR ≤1%) (Fig. 3B). To exclude the possibility of an association between the obtained results and the DBA/1 genotype, CA-I and S100A8 were assessed by WB in MP and PFP samples from C57BL/6 mice (n = 4 NI, n = 4 PbA-infected - ECM) (Supplementary Figure S5). CA-I was significantly more abundant in both MP and PFP samples from ECM mice compared to NI, while S100A8 was only detected in ECM MP, confirming the results obtained by both proteomics and WB in the DBA/1 model.
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