Figure 3.
MZB cell immunodepletion decreases anti-H2b antibody production in alloimmunized mice. (A) Representative dot plots of the gating strategy used for quantification of splenic MZB cells (B220+ IgM+ CD23− CD21++). BALB/C recipient mice were treated with anti-mouse CD11a and anti-mouse CD49d antibodies (lower panels) or their respective isotype-matched controls (IgG1 and IgG2a, upper panels). (B) Quantification of the percentage of splenic MZB cells among the CD45+ cells in mice treated with anti-CD11a/CD49d antibodies or their isotype controls (n = 5). (C) Anti-H2b IgG antibody production was evaluated weekly by flow cytometry in immunodepleted MZB cell (blue bars) and mice that received isotype control (red bars) BALB/C mice. Results are presented as the mean fluorescence intensity (MFI ± SEM) of Alexa-488 GaM IgG antibodies bound to H2b platelets (∗P < .05; ∗∗P < .01; 1-way ANOVA; n = 18). (D) The kinetics of elimination of transfused platelets were determined as the percentage of Oregon green-positive platelets among 10 000 RAM.1+ platelets in MZB cell depleted (blue line) and control mice (red line) at each time point (± SEM; ∗∗P < .01; ∗∗∗P < .001; 2-way ANOVA; n = 8-9). ANOVA, analysis of variance; SEM, standard error of the mean.

MZB cell immunodepletion decreases anti-H2b antibody production in alloimmunized mice. (A) Representative dot plots of the gating strategy used for quantification of splenic MZB cells (B220+ IgM+ CD23 CD21++). BALB/C recipient mice were treated with anti-mouse CD11a and anti-mouse CD49d antibodies (lower panels) or their respective isotype-matched controls (IgG1 and IgG2a, upper panels). (B) Quantification of the percentage of splenic MZB cells among the CD45+ cells in mice treated with anti-CD11a/CD49d antibodies or their isotype controls (n = 5). (C) Anti-H2b IgG antibody production was evaluated weekly by flow cytometry in immunodepleted MZB cell (blue bars) and mice that received isotype control (red bars) BALB/C mice. Results are presented as the mean fluorescence intensity (MFI ± SEM) of Alexa-488 GaM IgG antibodies bound to H2b platelets (∗P < .05; ∗∗P < .01; 1-way ANOVA; n = 18). (D) The kinetics of elimination of transfused platelets were determined as the percentage of Oregon green-positive platelets among 10 000 RAM.1+ platelets in MZB cell depleted (blue line) and control mice (red line) at each time point (± SEM; ∗∗P < .01; ∗∗∗P < .001; 2-way ANOVA; n = 8-9). ANOVA, analysis of variance; SEM, standard error of the mean.

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