Natural killer (NK) cells play a vital role in the rejection of tumors. Pancreatic ductal adenocarcinoma (PDAC), however, remains a poor prognosis malignancy, due to its resistance to radio‐ and chemotherapy, and low immunogenicity. We demonstrate here that IL‐2‐activated human NK cells are able to kill PDAC cells. Currently, novel strategies are being pursued to combat PDAC. In this regard, oncolytic viruses, in addition to killing tumor cells, may also have the potential to augment antitumor immune responses. We found that, besides having an intrinsic oncolytic activity, parvovirus H‐1PV is able to enhance NK cell‐mediated killing of PDAC cells. Our results show that H‐1PV infection of Panc‐1 cells increases NK cell capacity to release IFN‐γ, TNF‐α and MIP‐1α/β. Multiple activating receptors are involved in the NK cell‐mediated killing of Panc‐1 cells. Indeed, blocking of the natural cytotoxicity receptors—NKp30, 44 and 46 in combination, and NKG2D and DNAM1 alone inhibit the killing of Panc‐1 cells. Interestingly, H‐1PV infection of Panc‐1 cells overcomes the part of inhibitory effects suggesting that parvovirus may induce additional NK cell ligands on Panc‐1 cells. The enhanced sensitivity of H‐1PV‐infected PDAC cells to NK cell‐dependent killing could be traced back to the upregulation of the DNAM‐1 ligand, CD155 and to the downregulation of MHC class I expression. Our data suggests that NK cells display antitumor potential against PDAC and that H‐1PV‐based oncolytic immunotherapy could further boost NK cell‐mediated immune responses and help to develop a combinatorial therapeutic approach against PDAC.