Adenoviral vector has been extensively studied as a vaccine platform because of its ability to induce potent cellular and humoral immunity. One main advantage of adenoviral vectors is their natural tropism for mucosal surfaces, which makes them ideal for the purpose of mucosal vaccination against pathogens that preferentially initiate infection at the mucosal site. The current understanding of mucosal immunity suggests that mucosal vaccination is far superior to parenteral vaccination in protecting mucosal surfaces. Mucosal vaccination is particularly relevant to those infections for which parenteral immunization strategies have failed to confer protection. This review examines the use of adenoviral vector at mucosal sites for infectious disease against which the current vaccination strategies have been unsuccessful in eliciting protection. Data from animal models have suggested that adenoviral vectors are effective in protecting against infections caused by HIV, herpes simplex virus and Mycobacterium tuberculosis. We believe that these encouraging results will lead to further evaluation in clinical trials in the near future.