4―1BBL―B7―H3基因对免疫重建重症联合免疫缺陷荷瘤鼠抑瘤作用.docVIP

4―1BBL―B7―H3基因对免疫重建重症联合免疫缺陷荷瘤鼠抑瘤作用 　　[摘要] 目的 通过建立人免疫重建重症联合免疫缺陷（SCID）荷瘤鼠嵌合模型来探讨4-1BBL-B7-H3基因在非特异性抗肿瘤免疫中的作用。方法 将40只SCID鼠随机分为5组，A组（对照组）行免疫重建加注射Tca8113细胞；B组（Ad4-1BBL-B7-H3组）行免疫重建加注射含有人4-1BBL-B7-H3基因腺病毒转染的Tca8113细胞；C组（空载组）行免疫重建加注射含有空载体腺病毒转染的Tca8113细胞；D组（非免疫重建组）不给予小鼠免疫重建，注射Tca8113细胞；E组（非肿瘤组）行免疫重建加注射PBS。每周定期测量肿瘤体积；酶联免疫吸附法检测人IgG蛋白含量；流式细胞仪检测外周血中人CD3+、CD56+淋巴细胞比例；免疫组织化学法观察自然杀伤细胞2族成员D（NKG2D）和Toll样受体2（TLR2）在肿瘤中的表达；逆转录聚合酶链反应（RT-PCR）检测4-1BBL-B7-H3 mRNA的表达，实时定量聚合酶链反应（RT-qPCR）检测主要组织相容性复合体1类相关分子（M1C）A、B及TLR2的表达。结果 1）B组小鼠肿瘤体积最小（P0.05）；2）A、B、C、E组小鼠外周血中检测到人IgG、CD3+、CD56+淋巴细胞，B组淋巴细胞比例高于A、C和E组（P0.05）；3）B组NKG2D和TLR2的表达较其余各组明显增强；4）人4-1BBL-B7-H3基因在B组小鼠肿瘤中稳定表达；5）B组M1CA、M1CB和TLR2的表达高于A、C、D组（P0.05）。结论 4-1BBL-B7-H3基因在肿瘤组织中的高表达能成功诱导人CD3+、CD56+细胞增殖，直接或间接活化TLR2，上调NKG2D及其配体M1CA、M1CB的表达，从而产生有效的抗肿瘤免疫应答。 　　[关键词] 4-1BBL-B7-H3基因； 重症联合免疫缺陷鼠； 口腔鳞癌细胞株； 抗肿瘤免疫 　　[中图分类号] Q 78 [文献标志码] A [doi] 10.7518/hxkq.2014.02.019 　　[Abstract] Objective The non-specific antitumor immunity effect of 4-1BBL-B7-H3 gene was investigated by establishing an oral squamous cell carcinoma human peripheral blood lymphocyte-severe combined immunodeficient （SCID） mice chimeric model. Methods Forty mice were randomly divided into five groups. All groups， except the non-immune reconstitution group （group D）， had reconstructed human partial immune system. The control group （group A） was injected with Tca8113 cells. The Ad4-1BBL-B7-H3 group （group B） was injected with Tca8113 cells transfected by adenovirus containing 4-1BBL-B7-H3 gene. The empty vector group （group C） was injected with Tca8113 cells transfected by adenovirus containing an empty vector. The non-immune reconstitution group （group D） was injected with Tca8113 cells. The non-tumor group （group E） was injected with PBS. The tumor volumes in each group were measured weekly. Human IgG in blood was obtained through the tail vein and was determined by enzyme-linked immu-nosorbent assay. Human CD3+ and D56+ lymphocytes were assessed by flow cytometry. Model animals were kille