Assessment of humoral, innate, and T-cell immune responses to adeno-associated virus vectors.

Adeno-associated virus (AAV)-based gene therapy is being applied to treat a wide array of diseases. Pre-existing host immune responses to AAV and immune responses elicited by AAV vector administration remain a problem that needs to be further studied. Here, we present a series of protocols to assess immune responses before and after AAV vector administration that is applicable to multiple animal models and Phase I clinical trials. More specifically, to evaluate: 1) the humoral immune response, through levels of AAV-neutralizing and binding antibodies; 2) the innate immune response, through the acute induction of inflammatory cytokines; and 3) the T-cell immune response, through the activation of transgene- and vector-specific CD8 and CD4 T cells.