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. 2012 Sep;23(9):1538–1550. doi: 10.1681/ASN.2012020137

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Copyright © 2012 by the American Society of Nephrology

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Figure 10. — Antibody blockade of CD47 significantly ameliorates renal IRI. WT mice were treated with CD47 blocking antibody (αCD47) or isotype IgG control (CTRL) antibody (0.4 μg/g body weight) 90 minutes before induction of IRI. (A) Sham-operated kidneys were sectioned, stained with anti-rat AlexaFluor-555, and visualized by confocal microscopy. Representative images are shown. (B) Western blots of kidney tissue lysates were probed for TSP1 and CD47. Densitometry represents mean ± SD of n=4 samples per group. *P<0.001 WT + αCD47 antibody after IRI versus WT + CTRL antibody after IRI. (C) Levels of serum creatinine, and (D) quantitative analysis of tubular damage with representative renal tissue sections of corticomedulla from treatment groups 24 hours after ischemia-reperfusion are shown. Data are mean ± SD, n=10–12 per group. *P<0.001 and **P<0.0001 WT + αCD47 antibody after IRI versus WT + CTRL antibody after IRI. (E) Representative photomicrographs and quantitative analysis of kidney tissue sections stained by immunohistochemistry for neutrophils (total cell number per 10 hpf). Data shown are mean ±SD, n=6 per group and six independent fields assessed. (F) mRNA expression of proinflammatory cytokines IL-6, TNFα, and IL-1β, chemokines CCL2 and CXCL2, and iNOS in kidneys from WT mice pretreated with CTRL or αCD47 antibody and subjected to IRI. Results have been normalized to the housekeeping gene (HPRT1) and WT CTRL antibody animals used as the referent control. Data shown are mean ± SD, n=7 per group. **P<0.0001; ^∞P=0.90; ^#P=0.02. Original magnification, ×400 in A and E; ×200 in C.

Figure 10. — Antibody blockade of CD47 significantly ameliorates renal IRI. WT mice were treated with CD47 blocking antibody (αCD47) or isotype IgG control (CTRL) antibody (0.4 μg/g body weight) 90 minutes before induction of IRI. (A) Sham-operated kidneys were sectioned, stained with anti-rat AlexaFluor-555, and visualized by confocal microscopy. Representative images are shown. (B) Western blots of kidney tissue lysates were probed for TSP1 and CD47. Densitometry represents mean ± SD of n=4 samples per group. *P<0.001 WT + αCD47 antibody after IRI versus WT + CTRL antibody after IRI. (C) Levels of serum creatinine, and (D) quantitative analysis of tubular damage with representative renal tissue sections of corticomedulla from treatment groups 24 hours after ischemia-reperfusion are shown. Data are mean ± SD, n=10–12 per group. *P<0.001 and **P<0.0001 WT + αCD47 antibody after IRI versus WT + CTRL antibody after IRI. (E) Representative photomicrographs and quantitative analysis of kidney tissue sections stained by immunohistochemistry for neutrophils (total cell number per 10 hpf). Data shown are mean ±SD, n=6 per group and six independent fields assessed. (F) mRNA expression of proinflammatory cytokines IL-6, TNFα, and IL-1β, chemokines CCL2 and CXCL2, and iNOS in kidneys from WT mice pretreated with CTRL or αCD47 antibody and subjected to IRI. Results have been normalized to the housekeeping gene (HPRT1) and WT CTRL antibody animals used as the referent control. Data shown are mean ± SD, n=7 per group. **P<0.0001; ^∞P=0.90; ^#P=0.02. Original magnification, ×400 in A and E; ×200 in C.