Gene therapy is a clinical strategy that may potentially treat an array of genetic and nongenetic diseases, as well as a novel method for drug delivery and vaccination. To these ends, adenovirus vectors are a promising means to deliver specific genes of interest into the patient. A major limitation of the use of adenovirus vectors is the host immune response. Adenovirus vectors induce the innate arm of the immune system that results in inflammation of transduced tissues and efficient clearance of administered vectors. Unlike adaptive immunity, the innate response is mediated by the adenovirus particle and does not require viral transcription. In vivo, the innate immune response involves the induction of cytokines and activation of effector leukocytes that comprise the host response to these agents. A number of interactions with leukocytes and with epithelial and endothelial cells are essential in triggering the host response to adenovirus vectors. Signal transduction via MAP kinases and NF-κB-mediated gene transcription are triggered during early virus–cell interactions and are key events in the innate recognition of adenovirus vector transduction. This review aims to describe data examining cellular and molecular mechanisms involved in the adenovirus-mediated innate immune response.