The analysis of innate immune response to adenovirus using antibody arrays

Even though natural infections with adenovirus (Ad) are largely harmless in humans, an intravenous Ad vector administration for gene delivery purposes, especially at high doses, stimulates strong innate and adaptive immune responses, and can be fatal to the host. In animal models, intravenous Ad administration has been shown to induce transcription and release in the serum of a great number of pro-inflammatory cytokines and chemokines. Macrophages, including tissue residential macrophages (e.g., Kupffer cells in the liver), and dendritic cells throughout the body are considered to be the primary source of these pro-inflammatory mediators following their transduction with Ads. Here, we provide an overview and methodology for the qualitative and quantitative analyses of pro-inflammatory cytokine and chemokine expression in the spleen and their release into the bloodstream after intravenous Ad delivery using antibody arrays.