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. 2020 Sep 25;11:584254. doi: 10.3389/fimmu.2020.584254

FIGURE 5.

FIGURE 5

IFNα/β production and signaling promote K1 RBC alloimmunization. WT, IFNAR1–/–, and IRF3/7–/– mice were treated with pristane 14 days prior to analysis (A,B) or transfusion (C,D). Untreated WT mice were included as controls. (A) Quantification of Ly6G+ neutrophils, Ly6C+ monocytes, and CD11b+F4/80int inflammatory macrophages in peritoneal fluid; gated on Zombie-negative live cells. Inflammatory macrophages were also gated on TCRβ–B220– non-lymphocytes. (B) TNFα and IL-6 cytokine levels measured by multiplex array. Representative of 3 independent experiments with 5 mice per group. (C,D) Fourteen days after pristane-treatment, WT, IFNAR1–/–, and IRF3/7–/– mice were transfused with K1 RBCs. (C) Anti-KEL IgM was measured 4–5 days after transfusion. (D) Anti-KEL IgG data represent the peak IgG response 21–28 days after transfusion. 7–8 mice per group. Representative of 3 independent experiments. ∗p < 0.05 by Kruskal-Wallis test with a Dunn’s post-test.