Activation of DCs does not abrogate FIX tolerance induced by intramuscular injection of AAV1 in C57BL/6 mice. Plasma was collected every 4 weeks after AAV and LPS plus anti-CD40 antibody injection, and measured for hFIX antigen and anti-hFIX IgG antibodies by ELISA. Data shown are mean ± SEM, n = 5, for each cohort. (A) hFIX antigen. Eight- to 10-week-old C57BL/6 mice received intramuscular injection of 10¹¹ vg of AAV1-hFIX alone (■) or with LPS and anti-CD40 antibody (□). (B) Anti-hFIX IgG antibodies. Eight- to 10-week-old C57BL/6 mice received intramuscular injection of 10¹¹ vg of AAV1-hFIX alone (■) or with LPS and anti-CD40 antibody (□). (C) hFIX antigen. Eight- to 10-week-old C57BL/6 mice received intramuscular injection of 6 × 10¹⁰ vg of AAV2-hFIX alone (■) or with anti-CD40 antibody (□). (D) Anti-hFIX IgG antibodies. Eight- to 10-week-old C57BL/6 mice received intramuscular injection of 6 × 10¹⁰ vg of AAV2-hFIX alone (■) or with anti-CD40 antibody (□).