ppat.1009430.g004.tif (1.18 MB)
In vitro model of monocyte to macrophage differentiation during exposure to P. falciparum blood-stage parasites.
figure
posted on 2021-04-06, 17:24 authored by Rajan Guha, Anna Mathioudaki, Safiatou Doumbo, Didier Doumtabe, Jeff Skinner, Gunjan Arora, Shafiuddin Siddiqui, Shanping Li, Kassoum Kayentao, Aissata Ongoiba, Judith Zaugg, Boubacar Traore, Peter D. CromptonElutriated monocytes from healthy U.S. adults were incubated for 24 hours with medium alone, uninfected red blood cells (RBC) or Pf-iRBC (monocyte:Pf-iRBC ratio 1:15). At 24 hours, supernatants and cells were collected from some replicate wells for cytokine analysis, and the ChIP assay, while monocytes in other replicate wells were washed and incubated for 3 additional days in human serum plus medium to allow monocytes to differentiate into macrophages (Mf). On day 5, the three populations of macrophages (Mf, RBC-Mf and Pf-iRBC-Mf) were either harvested for ChIP and cytokine gene expression analysis; or re-stimulated with Pf-iRBCs or LPS for 24 hours prior to supernatants and cells being collected for cytokine analysis.
