Fig 2: CXCR4 chemokine receptor expression increase in the inflamed murine cornea. Representative confocal micrographs of 7-day suture inflamed BALB/c corneas show increased CD11c+ cells (red, A, D), CXCR4+ cells (green, B, E), and colocalization (yellow, C, F) throughout the corneal stroma of the peripheral (A–C) and central corneas (D–F). Orthogonal views highlight distribution throughout each corneal area. Cell density quantification of CD11c+ and CXCR4+ cells in peripheral and central corneal stroma (cells/mm2) highlights populations of CXCR4+ cDCs to be present following inflammation: significance for cell densities between central and peripheral cornea is denoted by asterisks (G). Representative histograms of flow cytometry analysis of relative CXCR4 expression within CD45+ leukocytes in the inflamed corneal epithelium (H) and stroma (I). Data are shown as mean ± SEM. t-test, **P < 0.01. Scale bars denote 50 µm. n = 4.

Fig 3: cDCs express CXCR4. Representative flow cytometry plots of CD11c+ cDCs from three spleens including cDC percentage of total splenocytes, expression of chemokine receptor CXCR4 in total cDCs and cDC subpopulations (CD8a+, CD11b+, and CD103+), and the cDC maturation marker MHC-II, respectively. CXCR4 was present in all CD8a+, CD103+, and CD11b+ cDC subsets, with similar expression in spleens from naïve BALB/c wild-type (A) and tumor mice (B). Analysis of CXCR4– splenic CD11c+ cDCs from wild-type (C) and tumor mice (D).

Fig 4: CXCR4 blockade decreases the recruitment of adoptively transferred cDCs. Pretreatment of adoptively transferred cDCs with anti-CXCR4–neutralizing monoclonal antibody reduces infiltration of adoptively transferred cDCs into the peripheral (A, B) and central (C, D) cornea 24 hours after injection compared with isotype control treatment, n = 7/group. Density quantification of cDC infiltration treated ex vivo with anti-CXCR4–blocking antibody reveal a significant decrease in the number of adoptively transferred cells compared with the appropriate isotype control (E) and total infiltration (F) into suture inflamed corneas. Scale bars denotes 50 µm. Data are shown as mean ± SEM. *P < 0.05, **P < 0.01. n = 4–7/group.