1. IL-27–eGFP is highly expressed by cDC1 cells and Ly6Chi monocytes after vaccine adjuvant administration.
IL-27–eGFP is highly expressed by cDC1 cells and Ly6Chi monocytes after vaccine adjuvant administration. Mice were immunized i.p. with combined polyIC/anti-CD40, and 6 h later spleens were harvested, digested in collagenase, and stained with Abs targeting markers for conventional DCs, monocytes, and granulocytes. (A and B) Using the gating strategies shown, eGFP expression was compared between unimmunized (red line graphs) and immunized (blue line graphs) reporter mice for cDC1 and cDC2 cells (A) or monocytes and granulocytes (B). The line graph marker was set based on the unimmunized sample, and the number represents the percentage of eGFP+ cells in the immunized sample. Quantification of the percentage of eGFP+ cells in each subset (C) and gMFI of eGFP in each subset (D). Error bars indicate SD derived from three mice each, in controls and immunized. (E) Spleens from mice immunized as in (A) were removed and fixed, and frozen sections were stained as described in Materials and Methods for B220 (blue), XCR1 (red), and eGFP (green). Original magnification ×22. Insets show magnified area marked by the yellow box. The experiment was performed three or four times, depending on the adjuvant. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < , t test.
Augustus M. Kilgore et al. ImmunoHorizons 2018;2:1-11
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