Here we report the effects of rotavirus infection on highly purified primary human pDCs. We demonstrate that although rotavirus initiates detectable transcription and translation in only a small percentage of pDCs, a significant percentage of pDCs activate and mature following exposure to virus. Importantly, we demonstrate that stimulation of pDCs with live or inactivated RRV effects secretion of IFNa and multiple proinflammatory cytokines and chemokines. This response is dependent on the presence of the viral dsRNA genome. As IFNa production by pDCs is classically triggered in response to ssRNA or DNA, the induction of this response by a replication-incompetent dsRNA virus indicates a potentially novel mechanism of viral sensing by pDCs.