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The Immological Rejection Activated by Human Heptocarcinoma Transferred with Murine H-2K b Gene
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Abstract:
To strengthen the immunogenicity of hepatocarcinoma cells and activate immnological cells recognizing and killing the tumor cells. Methods： The murine MHC-Ⅰ gene H-2K b which can express immunologial rejection antigens was transfected into human hepatocarcinoma cells HepG 2 by liposome DNA mediated gene transfer. The transfection and transcription of H-2K b gene were detected by molecular hybridization techniques. The exogeous antigens expressed on the membrane of transfected tumor cells were detected with ABC immunohistochemical method and flow cytometer. ［ 3H］ release assays were used to detect the recognizing and killing effects of lymphocytes to HepG 2 cells transferred with murine H-2K b gene. The nude mice experiment was used to further verify CTL cells killing active. Results: Southern blot hybridization showed that the H-2K b gene was integrated into the chromosome of HepG 2 cells. The RNA dot blot hybridization showed that there was transcription of H-2K b DNA in the transfected tumor cells. ABC immunohistochemical method and flow cytometer detection showed that the murine H-2K b antigens were expressed on the membrane of HepG 2 cells. ［3H］ release assays showed that the cytotoxicyty to HepG 2 cells transfected with H-2Kb gene was obviously higher than that to control cells. The results demonstrated that the growth of hepatocarcinoma cells which were transferred with H-2Kb gene was obviously inhibited. Conclusion： The murine MHC-Ⅰ gene H-2K b could be transferred into the human hepatocarcinoma cells and expressed on the membrane of transferred cells. The HepG 2 cells transferred with H-2K b gene could induce human effective lymphocytes to recognize and kill these transferred tumor cells.
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