4-1BBL-B7-H3基因对免疫重建重症联合免疫缺陷荷瘤鼠的抑瘤作用

发布时间：2018-04-23 14:20

本文选题：-BBL-B-H基因 + 重症联合免疫缺陷鼠　；参考：《华西口腔医学杂志》2014年02期

【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。
[Abstract]:Objective to investigate the role of 4-1BBL-B7-H3 gene in nonspecific anti-tumor immunity by establishing a chimeric model of human immunoreconstructive severe combined immunodeficient mice (SCID-bearing mice). Methods Forty SCID rats were randomly divided into five groups: group A (control group) receiving immune reconstitution plus Tca8113 cell group B (group B), and group C (no load group) treated with Tca8113 cells transfected with human 4-1BBL-B7-H3 gene adenovirus. Group D (non-immune reconstruction group) injected with empty vector adenovirus transfected Tca8113 cells did not receive immune reconstruction in mice. Tca8113 cells were injected into E group (non tumor group). The tumor volume was measured weekly, the content of human IgG protein was detected by enzyme-linked immunosorbent assay (Elisa), and the proportion of human CD3 CD56 lymphocytes in peripheral blood was detected by flow cytometry. Immunohistochemical method was used to observe the expression of DnkG2D) and Toll like receptor 2 (TLR2) in tumor, and reverse transcriptase polymerase chain reaction (RT PCR) was used to detect the expression of 4-1BBL-B7-H3 mRNA. Real-time quantitative polymerase chain reaction (RT-PCR) was used to detect the expression of major histocompatibility complex class 1 related molecule, M1CnAmb and TLR2. Results 1 the percentage of NKG2D and TLR2 in the peripheral blood of the mice in group B was higher than that in group A C and group E, and the expression of human 4-1BBL-B7-H3 gene in group B was significantly higher than that in group B (P 0.05). The expression of 4-1BBL-B7-H3 gene in group B was significantly higher than that in group A (P 0.05) and the expression of 4-1BBL-B7-H3 gene in group B was significantly higher than that in group B (P < 0.05), and the expression of 4-1BBL-B7-H3 gene in group B was significantly higher than that in group B (P < 0.05). The expression of M1CAN M1CB and TLR2 in group B was higher than that in group A (P 0.05). Conclusion the high expression of 4-1BBL-B7-H3 gene in tumor tissue can successfully induce the proliferation of human CD3 cell line CD56, activate TLR2 directly or indirectly, and up-regulate the expression of NKG2D and its ligand M1CAAM1CB, thus producing an effective anti-tumor immune response.
【作者单位】：北京大学深圳医院口腔颌面外科;汕头大学医学院;
【基金】：国家自然科学基金资助项目(30973339) 广东省自然科学基金资助项目(S2012010010382) 广东省科技厅国际合作基金资助项目(2010B050100007) 深圳市基础研究基金资助项目(JC2011-05201030A) 深圳市国际合作基金资助项目(ZYA201106100080A)
【分类号】：R739.8

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