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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IFNγ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20mg/L;并对OC1669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IFNγ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。
OBJECTIVE: To study the antitumor immunity of 6B11 anti-idiotypic antibody in vitro and explore its potential as a vaccine for ovarian cancer. Methods: Peripheral blood mononuclear cells were isolated and stimulated with 6B11 anti-idiotypic antibody and cultured. The 3HTdR incorporation method and 51Cr cytotoxicity were used to observe the proliferation and cytotoxicity of lymphocytes respectively. The levels of IFNγ in culture supernatants stimulated with 6B11 micro-antibody were measured by ELISA. The expression of IFNγ in lymphocytes was detected by flow cytometry Phenotypic changes. Results: The anti-idiotypic antibody 6B11 could stimulate the proliferation of human peripheral blood lymphocytes with the best stimulation dose of 20mg / L. The anti-idiotypic antibody 6B11 had the cytotoxic effect on ovarian cancer cell lines with OC1669 expression. The level of IFNγ in the supernatant increased. The phenotypes of lymphocytes showed that the number of CD3 + cells was significantly increased, the number of CD4 + cells was increased and the number of CD8 + cells was not increased obviously. CD4 + / CD8 + ratio was significantly higher, the difference was statistically significant. CONCLUSION: 6B11 anti-idiotype micro-antibody can induce specific humoral and cellular immune responses in vitro and provide experimental evidence for the clinical application of anti-idiotype micro-antibody vaccine.