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H22细胞全细胞抗原负载的树突状细胞激活肿瘤浸润性淋巴细胞抗小鼠肝癌的实验

Anti-mouse Hepatoma Effect of Tumor-infiltrating Lymphocyte Stimulated by DCs Pulsed with H22 Full-cell Antigen

  • 摘要: 目的 探讨H22小鼠肝癌细胞(H22细胞)全细胞抗原致敏的树突状细胞激活肿瘤浸润淋巴细胞抗小鼠肝癌细胞活性。方法取得小鼠骨髓细胞并诱导生成树突状细胞,由冻融法制备的H22细胞全细胞抗原致敏,然后用已致敏的树突状细胞激活肿瘤浸润性淋巴细胞,测定致敏前后的DC表面抗原CD11c、CD80、CD86、CD40、MHCⅡ,并评估激活前后的TIL对H22细胞的杀伤活性,同时脾淋巴细胞作为杀伤对照。结果CD11c阳性细胞中CD80、CD86、CD40、MHCⅡ阳性细胞所占比例在致敏后的DC表现为明显上调。经致敏后成熟DC激活的TIL对H22细胞杀伤活性明显高于未激活的TIL,并高于激活或未激活的小鼠脾脏淋巴细胞。结论在H22细胞全抗原致敏后,小鼠成熟DC中CD80、CD86、CD40、MHCⅡ的表达率明显高于未成熟DC。经H22细胞全细胞抗原致敏的DC能诱导活化TIL,明显提高其在体外对H22细胞的杀伤活性。

     

    Abstract: Objective To study the anti-mouse hepatoma effect of tumor-infiltrating lymphocytes (TILs) stimulated by dendritic cells (DCs) pulsed with the tumor full-cell antigen in vitro. Methods DCs were isolated from BALB/c mouse bone marrow and stimulated by granulocyte macrophage colony stimulating factor (GM-CSF),interleukin-4(IL-4) and H22 full-cell antigen.The maturation surface markers CD80,CD86,CD40,MHCⅡ and CD11c were detected by flow cytometry before and after pulsing with whole H22 cells lysates.Then the cytotoxic potency of the TILs was assessed both stimulated and unstimulated,and taking the spleen lymphocytes as control groups.ResultThe expression of CD80,CD86,CD40,MHCⅡon the DCs pulsed with H22 full-cell lysate is a higher than the DCs after 5d of culture.TILs stimulated by the DCs on mouse hepatoma H22 cells had very special efficient anti-tumor effect on the mouse liver cancer H22 in vitro.And the anti-tumor effect of stimulated TIL is higher than the TILs and spleen lymphocytes. Conclusion The results suggest that H22 tumor full-cell antigens specific induced tumor-infiltrating lymphocytes have more efficient at killing the H22 cells.

     

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