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Specific antitumorimmunity induced by mIL12 gene modified dendritic cells transfected with murine colon carcinoma RNA
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Abstract:
To investigate the specific antitumor effect of mIL12 gene modified dendritic cells (DC) transfected with the total RNA of CT26 murine colon carcinoma. Methods: DC were cultured from murine bone marrow in the presence of rmGMCSF and rmIL4.The purity of DC was detected by flowcytometry. Adenovirus carrying mIL12 gene was proliferated in 293 cells and was transfected into DC. The total RNA of CT26 was extracted by Trizol reagent and was introduced into mIL12 gene modified DC by TransMessenger in vitro. The modified DC were used to immunize mice. The in vitro and in vivo levels of mIL12 and the in vivo activity of cytotoxic T lymphocyte（CTL）were examined by ELISA assay and modified LDH release assay, respectively. The tumor growth and animal survival time of immunized mice were estimated after they were challenged with parental tumor cells. Results: DC were successfully obtained from the culture of murine bone marrow, with CD11c+ accounting for over 90%. Vaccination with mIL12 gene modified DC transfected with the total RNA of CT26 induced strong specific CTL activity in vitro. All the immunized mice survived for a long term. Vaccination with AdLacZ modified DC and DC transfected with the total RNA induced moderate specific CTL activity in vitro（P<0.01）, and 60% of the immunized mice survived for a long time. There was no specific CTL activity in the mice immunized with DC and PBS and all the mice died. Conclusion: mIL12 gene modified DC transfected with the total RNA of CT26 can effectively present endogenetic tumor antigen and induce strong CTL activity, thus inducing more specific antitumor immune response.
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Clc Number:
R730.3
