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Virotherapy in one of the main current applications of recombinant adenoviruses. Oncolytic adenovirus are designed to target tumors, replicate selectively in tumor cells, and elicit immune responses against tumor antigens. Transgene expression in replication-competent oncolytic vectors allows to explore multiple strategies to enhance the potential of virotherapy. In this chapter we describe common in vivo and in vitro techniques used to evaluate the potency and biodistribution of oncolytic viruses. Monitoring immune responses against viral and tumor antigens is crucial as the immune system determines the outcome of virotherapy.