卵巢癌6B11抗独特型微抗体诱导抗肿瘤免疫应答的体外实验

目的:研究6B11抗独特型微抗体在体外抗肿瘤免疫情况,探讨其作为卵巢癌疫苗的可能性.方法:分离人外周血单个核细胞,用6B11抗独特型微抗体刺激并培养.采用3HTdR参入法、51Cr细胞毒实验分别观察淋巴细胞增殖情况和细胞毒作用,ELISA方法测定用6B11微抗体刺激后培养上清液中IFN-γ水平的变化,流式细胞术检测微抗体刺激后淋巴细胞表型的改变.结果:6B11抗独特型微抗体可以刺激人外周血淋巴细胞的增殖,最佳刺激剂量为20 mg/L;并对OC166-9表达阳性的卵巢癌细胞系有细胞毒作用;用6B11微抗体刺激后培养上清液中IFN-γ水平升高;淋巴细胞的表型表现为,疫苗刺激后CD3 的细胞明显增高, CD4 的细胞也增加, CD8 的细胞增加不明显.CD4 /CD8 的比率明显增高,差异有统计学意义.结论:6B11抗独特型微抗体在体外可诱导机体产生特异体液免疫和细胞免疫反应,为抗独特型微抗体疫苗的临床应用提供了一定的实验依据.

In vitro study of the antitumor immune responses induced by anti-idiotypic minibody vaccine of ovarian cancer

OBJECTIVE: To evaluate whether anti-tumor immune response can be induced in vitro with 6B11 anti-idiotypic minibody. and to explore its probability as ovarian cancer vaccine. METHODS: Separated human peripheral blood mononuclear cells (PBMC) were stimulated and cultured by 6B11 minibody. The proliferations of PBMC and cytotoxin were observed by (3)HTdR and (51)Cr release test respectively. ELISA(Enzyme-Linked Immunosorbent Assay) test and Immune Flow Cytometry were used to analyze IFN-gamma in supernatant of the cultured cdlls and the change of T lymphocyte phenotype of PBMC with 6B11 minibody stimulated. RESULTS: 6B11 minibody could stimulate PBMC to proliferate, the best dose was 20 mg/L; it performed cytotoxin function to ovarian carcinoma cell line expressing OC166-9. IFN-gamma maintained at high level after stimulation. It stimulated proliferation of CD3(+) T cell and CD4(+) from PBMC after stimulation respectively. CD8(+) T cell proliferation was not clear. There was significant difference between stimulation and unstimulation in CD4(+)/CD8(+) ratio. CONCLUSION: 6B11 anti-idiotypic minibody can induce both humoral and cellular immunity against ovarian carcinoma in vitro. This paper has provided strong experimental evidence for clinical use of 6B11 minibody as anti-idiotype vaccines against ovarian carcinoma.