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. 2014 Sep 29;26(4):896–906. doi: 10.1681/ASN.2014020195

Figure 5.

Figure 5.

Upregulated ICAM-1 expression in the absence of CD169+ cells exacerbates AKI. (A) Chemokine expression in IR kidneys. DT was injected intraperitoneally into WT or CD169-DTR mice. Thirty-six hours later, bilateral IR was performed. Quantities of ccl2, cxcl1, and cxcl2 mRNA levels in the kidneys relative to WT preinjury mice were determined by quantitative PCR. n=5 mice/group. Mean±SD. *P<0.05. (B) Enhanced expression of ICAM-1 in kidneys of CD169-DTR mice. Quantity of icam1 and icam2 mRNA levels in the kidneys of DT-treated WT or CD169-DTR mice relative to WT mice without DT were determined by quantitative RT-PCR. n=8 mice/each group. The average values are plotted with SD. *P<0.05. (C) Blockade of ICAM-1 suppresses the development of AKI in CD169-DTR mice. DT was injected intraperitoneally into CD169-DTR mice. Twenty-four hours later, 100 μg anti–ICAM-1 (clone KAT1) antibody or PBS was injected intravenously into these mice. Thirty-six hours after the DT injection, bilateral IR was performed. Survival rate is shown. n=3 mice/group. *P<0.05. (D) CD169+ cells localize in the perivascular area of the renal interstitium. DyLight 488-labeled tomato lectin was injected intravenously into CD169-Cre-YFP mice 5 minutes before removing the kidney. Cryosections of the kidneys were stained for YFP (green) and observed under a confocal microscope. White arrows indicate the CD169-positive macrophages, which are located along the capillary vessels. z-Stack images were processed by ImageJ software. Red signal indicates tomato lectin on endothelium. Scale bar, 100 μm. (E) Peripheral blood leukocytes from CD169-DTR mice augment ICAM-1 expression levels in cultured endothelial cells. (Left panel) Peripheral blood leukocytes from DT-treated WT or CD169-DTR mice were cocultured with UV♀2 cells for 18 hours. icam1 mRNA levels of these cells were determined by quantitative RT-PCR. (Center panel) Peripheral blood leukocytes from DT-treated CD169-DTR mice were cocultured with UV♀2 cells for 18 hours in the presence or absence of butylated hydroxyanisole (BHA). (Right panel) Peripheral blood leukocytes from DT-treated WT or DT-treated CD169-DTR mice were cocultured with UV♀2 cells for 18 hours using the transwell plate. The average values are plotted with SD. Data are representative of two or three independent experiments. *P<0.05.