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. 2015 Dec 1;4(12):e265. doi: 10.1038/mtna.2015.40

Figure 3.

Figure 3

In vivo administration of a CD40 antisense oligonucleotide (ASO) resulted in robust reduction of kidney CD40 and CD40-dependent inflammation. (a,b) To evaluate CD40 ASO activity following a broad induction of inflammation, C57BL/6 mice were given 40 mg/kg/week of ASO treatments for 4 weeks followed by 0.5 mg/kg lipopolysaccharide (LPS) given 4 hours before kidneys were harvested. (a) Kidney CD40 and (b) CCL5 expression as measured by quantitative real-time polymerase chain reaction (qRT-PCR). (c–e) To evaluate CD40 ASO activity following CD40-dependent inflammation, C57BL/6 mice were given 40 mg/kg ASO treatments on days 0 and 7, followed by 25 μg of the activating CD40 mAb on day 10 and then kidneys were harvested on day 11 and (c) kidney CD40 was measured by enzyme-linked immunosorbent assay (ELISA) and qRT-PCR, (d) ICAM1, VCAM1, E Selectin, CCL5 and IL12p40 were measured by qRT-PCR, and (e) kidney CCL5 was measured by ELISA. (f) WT and CD40 KO recipient mice were irradiated with 7 Gy and then given IV ~400K bone marrow (BM) donor cells of the specific genotype. Seven weeks following reconstitution, animals received two SC administrations of 40 mg/kg ASO (study days 0 and 7) and then they received 25 μg of the activating CD40 mAb (on day 10). Kidneys were harvested on day 11 for mRNA analysis of CD40 and CD40-dependent inflammation. For all datasets, kidney CD40 or CCL5 protein was measured by ELISA and mRNA expression measured by qRT-PCR. Data are reported as relative expression to (+) CD40 Ab, phosphate buffered saline (PBS) group = 1.0, unless otherwise noted. Mean ± SEM, n = 4–6/group, *P < 0.05 versus (+) CD40 Ab, PBS group (a–e) or versus (+) CD40 Ab, (−) CD40 ASO, WT/BM:WT (f).