Figure 5
Figure 5. FOXP1 overexpression specifically inhibits formation of IgG- but not IgM-secreting PCs. (A-B) CD19+CD27+ MBCs were sorted from human peripheral blood and transduced with FOXP1-IRES-YFP, BCL6-IRES-GFP, or control empty vector and cultured under conditions that promote PC differentiation. Six days after transduction, YFP- or GFP-positive cells of cultures transduced with FOXP1, BCL6, or control vector were sorted. (A) Equal numbers of sorted cells (50 000) were cultured with IL-21 and IL-2 for an additional 24 hours. Thereafter, the supernatants were collected, and IgM and IgG protein levels were analyzed by ELISA. Levels were normalized to levels in control-transduced cells. Means ± SD of at least 4 independent experiments are shown. (B) Equal numbers of sorted cells were plated onto membranes in serial dilutions and cultured with IL-21 and IL-2 for an additional 18 hours, after which numbers of IgM- or IgG-secreting cells were determined by ELISPOT. Spot numbers were normalized to numbers in stimulated, control-transduced cells. Means ± SD of 3 independent experiments are shown (1 sample t test, *P < .05; **P < .01; ***P < .001).

FOXP1 overexpression specifically inhibits formation of IgG- but not IgM-secreting PCs. (A-B) CD19+CD27+ MBCs were sorted from human peripheral blood and transduced with FOXP1-IRES-YFP, BCL6-IRES-GFP, or control empty vector and cultured under conditions that promote PC differentiation. Six days after transduction, YFP- or GFP-positive cells of cultures transduced with FOXP1, BCL6, or control vector were sorted. (A) Equal numbers of sorted cells (50 000) were cultured with IL-21 and IL-2 for an additional 24 hours. Thereafter, the supernatants were collected, and IgM and IgG protein levels were analyzed by ELISA. Levels were normalized to levels in control-transduced cells. Means ± SD of at least 4 independent experiments are shown. (B) Equal numbers of sorted cells were plated onto membranes in serial dilutions and cultured with IL-21 and IL-2 for an additional 18 hours, after which numbers of IgM- or IgG-secreting cells were determined by ELISPOT. Spot numbers were normalized to numbers in stimulated, control-transduced cells. Means ± SD of 3 independent experiments are shown (1 sample t test, *P < .05; **P < .01; ***P < .001).

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