Activation of the NF-kappa B pathway by adeno-associated virus (AAV) vectors and its implications in immune response and gene therapy

作者：Jayandharan Giridhara R; Aslanidi George; Martino Ashley T; Jahn Stephan C; Perrin George Q; Herzog Roland W; Srivastava Arun^*

来源：Proceedings of the National Academy of Sciences of the United States of America, 2011, 108(9): 3743-3748.

DOI：10.1073/pnas.1012753108

Because our in silico analysis with a human transcription factor database demonstrated the presence of several binding sites for NF-kappa B, a central regulator of cellular immune and inflammatory responses, in the adeno-associated virus (AAV) genome, we investigated whether AAV uses NF-kappa B during its life cycle. We used small molecule modulators of NF-kappa B in HeLa cells transduced with recombinant AAV vectors. VP16, an NF-kappa B activator, augmented AAV vector-mediated transgene expression up to 25-fold. Of the two NF-kappa B inhibitors, Bay11, which blocks both the canonical and the alternative NF-kappa B pathways, totally ablated transgene expression, whereas pyrrolidone dithiocarbamate, which interferes with the classical NF-kappa B pathway, had no effect. Western blot analyses confirmed the abundance of the nuclear p52 protein component of the alternative NF-kappa B pathway in the presence of VP16, which was ablated by Bay11, suggesting that AAV transduction activates the alternative NF-kappa B pathway. In vivo, hepatic AAV gene transfer activated the canonical NF-kappa B pathway within 2 h, resulting in expression of proinflammatory cytokines and chemokines (likely reflecting the sensing of viral particles by antigen-presenting cells), whereas the alternative pathway was activated by 9 h. Bay11 effectively blocked activation of both pathways without interfering with long-term transgene expression while eliminating proinflammatory cytokine expression. These studies suggest that transient immunosuppression with NF-kappa B inhibitors before transduction with AAV vectors should lead to a dampened immune response, which has significant implications in the optimal use of AAV vectors in human gene therapy.

• 出版日期2011-3-1