Figure 3
Elevated IL-10 is essential for tolerance to CII. (A) Mice (n = 3/group) were injected intravenously with 1011 VG of AAV8.CII. At day 14 mice were injected intravenously with 1010 VP Ad5.CII. 10 days after Ad5.CII injection liver non-parenchymal cells were cultured with anti-CD3 and anti-CD28 in the presence of CII; 96-h later culture supernatants were collected, pooled and analyzed for cytokine production. Dot blot analysis of cytokine production from cultured liver non-parenchymal cells stimulated with CII. Pixel density was measured with ImageJ software (version 1.51; https://imagej.nih.gov/ij/). (B) Quantification of IL-10 production in cell culture supernatants was analyzed by ELISA. Mice were treated as in A except that 0.75 mg of PC61 anti-CD25 antibody was injected in some mice 7 days after AAV8.CII administration (C) 10 days after Ad5.CII injection, liver non-parenchymal cells were stimulated with CII and IFNγ production was measured by ELISPOT. Spots were quantified using the ISPOT software (version 7, https://www.elispot.com/products/software/). Mice were treated as in A except that one group of mice which received 0.75 mg of 1B1.3A anti-IL-10R antibody intraperitoneally 2 days prior and 2 days post AAV8.CII injection. (A–C) For each group, the mean ± SEM is shown.
