Figure 4
AAV8.CII prophylaxis reduces CIA severity through the induction of Tregs and IL-10 production. (A–H) Mice (n = 5/group) were injected intravenously with 1011 VG of AAV8.CII. 14 days later they received 100 ug CII emulsified in 100 ug CFA by subcutaneous injection. 21 days later the mice were injected intraperitoneally with 50 ug of LPS suspended in PBS. Peripheral inflammation was followed for the next 15 days and prior to euthanasia. (A) Experimental timeline is illustrated (B) Serum taken 15 days post LPS injection was assessed for anti-CII IgG by ELISA. (C) 15 days post LPS injection, liver non-parenchymal cells were isolated and stained with CD4, CD25 and FoxP3 antibodies run by FACS and the percentage of triple positive cells were compared using FlowJo, version 10.7 (www.flowjo.com). (D) AAV8.CII was injected and disease induction performed as per (A) with an additional administration of PC61 on day 2. (E) Peripheral inflammation was measured in the paws of arthritis mice every 2–3 days. Data presented is the summation of the four paws to a maximum inflammation score of 16 per day. (F) Serum taken 15 days post LPS injection was assessed for anti-CII IgG by ELISA. (G&H) IFNγ and IL-10 production in cell supernatant was assessed as in 3.D. by ELISA. (B,C,F–H) Means ± SEM are depicted for the various assays.
