Journal of Immunology Research / 2019 / Article / Fig 2

Research Article

Endothelial Protein C Receptor Could Contribute to Experimental Malaria-Associated Acute Respiratory Distress Syndrome

Figure 2

TNF increases Plasmodium berghei adherence in the ARDS experimental model. (a) Serum from ARDS- and HP-developing mice was collected on the 7th day postinfection (dpi) and submitted for TNF analysis by ELISA. (b) Primary microvascular lung endothelial cells (PMLECs) from naïve DBA/2 mice were stimulated with infected red blood cells (iRBCs), and TNF was measured in the supernatant. (c–e) PMLECs were also stimulated with TNF (50 ng/ml), and the adhesion of iRBCs to PMLECs under static conditions and (f–h) under flow conditions (after 24 hours of TNF stimulation) was analyzed. (i–k) iRBCs adhered in PMLECs in a static condition assay after coincubation with peritoneal macrophages (Mφ) seeded in transwell membranes and coincubated with iRBCs or red blood cells (RBCs) for 24 hours, and then (j) TNF release was measured in the supernatant. (k) PMLECs (that had contact with the Mφ supernatant) were incubated with iRBCs having their adhesion analyzed. (l) iRBC adherence was assessed in TNF-stimulated cells and TNF-neutralized antibodies plus TNF. Bars represent the . Graphs: (a) one-way ANOVA from two grouped experiments; (b–h) the Mann-Whitney test is representative of (b and h) two independent experiments and (c–e and l) two grouped experiments; (j and k) the Kruskal-Wallis test is representative of two independent experiments; bars represent the (; ; ). ARDS: acute respiratory distress syndrome; HP: hyperparasitemia. Arrows in (f), (g), and (i) indicates iRBC adhered in PMLECs.

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