Wu L.-H.,Tumor Research Institute of Hebei Province |
Zhang C.,Tumor Research Institute of Hebei Province |
Shan B.-E.,Tumor Research Institute of Hebei Province |
Liu L.-H.,Tumor Research Institute of Hebei Province |
Ai J.,Tumor Research Institute of Hebei Province
Chinese Journal of Cancer Biotherapy | Year: 2010
Objective: To explore the in vitro and in vivo effects of CD40L-expressing murine colon cancer colon26 vaccine on activities of dendritic cells (DCs). Methods: The CD40L gene was transfected into colon26 cells using lipofactamine method, and the stable colon26/CD40L transfectants were obtained. Colon26/CD40L cells were co-cultured with DCs, and the phenotype of DCs was examined by flow cytometry; the expressions of cytokine genes such as IL-12, IL-23, IL-18 and IFN-γ were examined by RT-PCR; and the IL-12, IL-23 and IFN-γ levels in co-cultured supernatants were measured by ELISA. Tumor-bearing BALB/c mouse model was prepared by subcutaneous injection of colon26 cells, and treated with DCs impulsed with colon26/CD40L vaccine. IL-12, IL-23, IFN-γ, IL-10 and TGF-β levels in mouse peripheral blood were detected by ELISA; and the histopathological changes of the liver, spleen and tumor tissues were observed by H-E staining. Results: Colon26 cells stably expressing CD40L (colon26/CD40L vaccine) were successfully obtained. DCs, when co-cultured with colon26/CD40L vaccine, had up-regulated expressions of co-stimulatory molecules, including CD80, CD86, MHC I and MHC (II (P < 0.01). The expressions of IL-12, IL-23, IL-18 and IFN-γ mRNA genes were only detected in DC-colon26/CD40L system, not in other co-cultured systems. And the DC-colon26/CD40L co-cultured system showed higher IL-12, IL-23 and IFN-γ levels (P < 0.01). Compared with mice treated with colon26/CD40L or DCs, mice treated with DCs-colon26/CD40L had smaller tumor volumes and lower weights (P < 0.05), increased IL-12, IL-23, and IFN-γ levels in the peripheral blood (P < 0.01), decreased IL-10 and TGF-β levels (P < 0.01), and better histopathological changes of tumor tissues. Conclusion: CD40L-expressing colon cancer cells can promote the maturation, induce the cytokine secretion, and enhance the activities of DCs in vitro and in vivo.