Journal: PLoS Neglected Tropical Diseases
Article Title: Leishmania infantum Amastigotes Trigger a Subpopulation of Human B Cells with an Immunoregulatory Phenotype
Figure Lengend Snippet: L . infantum amastigotes activate human B cells. Purified human B cells were either left alone (used as a control) or incubated overnight with L . infantum amastigotes (AMA) at a final parasite:host cell ratio of 3:1. Cells and the cell-parasite mixture were intensively washed with a galactose-modified PBS/EDTA solution and stained with the listed antibodies. Samples were read using a BD FACSCanto flow cytometer. A) Representative histograms displaying expression of each cell surface marker studied are shown in this panel. Dark lines on the histograms represent the fluorescence minus one (FMO) control for each sample. B) Results represent mean values of samples from 5 to 7 different healthy donors and are expressed as the percentages of positive cells for the indicated cell surface marker. C) Results represent mean values of samples from 5 to 7 different healthy donors and are expressed as the mean fluorescence intensities (MFI) for the indicated cell surface marker. P values were calculated by the two-tailed Student’s t-test (ns: not significant).
Article Snippet: Viability of B cells was assessed by staining with the Fixable Viability Dye eFluor 780 in PBS/EDTA 2 mM, following manufacturer’s instructions (eBiosciences, San Diego, CA).
Techniques: Purification, Incubation, Modification, Staining, Flow Cytometry, Cytometry, Expressing, Marker, Fluorescence, Two Tailed Test