23Introduction. Platelets drive endothelial cell activation in many diseases. 24 However, if this occurs in Plasmodium vivax malaria is unclear. As platelets have 25 been reported to be activated and to play a role in inflammatory response during 26 malaria, we hypothesized that this would correlate with endothelial alterations 27 during acute illness.
28Methods. We performed platelet flow cytometry of PAC-1 and P-selectin. We 29 measured Platelet markers (CXCL4, CD40L, P-selectin, Thrombopoietin, IL-11) and 30 endothelial markers (ICAM-1, von Willebrand Factor and E-selectin) in plasma with a 31 multiplex-based assay. The values of each mediator were used to generate heatmaps, K-32 means clustering and Principal Component analysis. In addition, we determined pair-wise 33 Pearson's correlation coefficients to generate correlation networks. 34 Results. Platelet counts were reduced, and mean platelet volume increased in 35 malaria patients. The activation of circulating platelets in flow cytometry did not differ 36 between patients and controls. CD40L levels (Median [IQ]: 517 [406-651] vs. 1029 [732-37 1267] pg/mL, P=0.0001) were significantly higher in patients, while P-selectin (Median 38 [IQ]: 17.0 [15.4-20.6] vs. 22.2 [17.6-25.7] ng/mL, P=0.0621) and CXCL4 showed a 39 nonsignificant trend towards higher levels in patients. The network correlation 40 approach demonstrated the correlation between markers of platelet and endothelial 41 activation, and the heatmaps revealed a distinct pattern of activation in two subsets 42 of P. vivax patients when compared to controls.43 Conclusion. platelet activation occurs in uncomplicated vivax malaria and 44 this correlates with higher endothelial cell activation, especially in a subset of 45 patients. 46 47 131 (R&D Systems): CD40L, P-selectin, PF4 and thrombopoietin (TPO), IL-11, as 132 well as circulating markers of EC activation (ICAM-1, E-selectin, von Willebrand 133Factor (vWF)). We selected 31 patients for the multiplex assay from a wide range 134 of parasitemias (260 to 25,150 Pv-IE/μL), so that a wide spectrum of disease 135 burden could be evaluated. This subgroup did not differ from the overall 136 population of patients regarding severity of disease, sex proportion, platelet counts 137 and mean parasitemia. We selected nine controls matched for age and sex. For the 138 network analysis, we used some interpolated results from below standard range. 139 140