Pancreatic cancer remains a lethal malignancy. Despite chemotherapy or/and radiotherapy after the surgery, the improvement on the overall survival of the patients has still been minimal. To develop novel therapeutic approaches, we tried to prepare mHSP65-TTL, a candidate vaccine prepared by mixing the recombinant mycobacterial heat shock protein 65 (mHSP65) with tumor tissue lysate (TTL) of Panc02 pancreatic cancer tissue. The mHSP65-TTL were used to immune the C57BL/6 mice implanted with the Panc02 cancer cells, in combination with or without low dose cyclophosphamide (CY). The results showed that mHSP65-ITL significantly prolonged the survival of the pancreatic cancer bearing mice and low dose CY enhanced the efficacy of the mHSP65-TTL. In addition, we detected mRNA expression of ROR gamma t and IL-17A in spleen cells of mice received mHSP65-ITL or mHSP65-TTL plus CY, and found that mHSP65-TTL up-regulated mRNA expressions of ROR gamma t and IL-17A, CY alone or mHSP65-TTL plus CY up-regulated mRNA expressions of ROR gamma t. The work could provide an insight into a combinational approach for the immunotherapy of pancreatic cancer. (C) 2015 Published by Elsevier B.V.