论文部分内容阅读
目的: 分析抗CD3分子的单克隆抗体 (mAb)yCD3的免疫学特性和生物学活性, 观察其激活的免疫活性细胞CD3AK体外及动物体内的抑瘤作用。方法: 用流式细胞术(FCM)测定yCD3的特异性, 以及CD3AK细胞的免疫表型和产生细胞因子的情况。用 3H -TdR法测定yCD3对淋巴细胞转化的作用; 乳酸脱氢酶法 (LDH)测定CD3AK细胞的体外细胞毒活性。建立荷瘤小鼠模型, 观察静脉注射CD3AK细胞后肿瘤生长的情况、转移灶数量和小鼠存活天数。结果: yCD3与T细胞呈特异性反应, 5μgyCD3可竞争抑制 70%的标准抗CD3抗体与细胞表面CD3分子的结合。yCD3刺激外周血淋巴细胞增殖的有效浓度为 8μg/L, 并与IL- 2、抗CD28抗体有协同作用。活化的CD3AK细胞中CD3+、CD8+和CD25+细胞增多; 产生IL- 2和IFN γ的CD3+细胞均有不同程度的增加, 在抗CD28抗体协同刺激下分别增加 3. 29和 2. 47倍。当效靶细胞比为 80∶1时, CD3AK细胞对体外肿瘤细胞杀伤的百分率为 57. 54%。分组观察荷瘤动物, CD3AK细胞治疗组的抑瘤率为 33. 17%, 对小鼠肿瘤肺转移的抑制率为39. 70%, 与LAK细胞联合治疗的疗效更显著。结论: yCD3可活化T细胞, 诱导的CD3AK细胞在体外及动物体内显示抑制肿瘤的作用, 在临床抗肿瘤过继性免疫治疗中具有重要意义。
OBJECTIVE: To analyze the immunological characteristics and biological activity of anti-CD3 monoclonal antibody (mAb) yCD3 and to observe the anti-tumor effect of CD3AK activated in vitro and in vivo by its activated immunocompetent cells. Methods: The specificity of yCD3 was determined by flow cytometry (FCM) and the immunophenotype and cytokine production of CD3AK cells. The effect of yCD3 on lymphocyte transformation was determined by 3H-TdR method. The cytotoxic activity of CD3AK cells was measured by lactate dehydrogenase (LDH) assay. The tumor-bearing mice model was established, the growth of tumor after intravenous injection of CD3AK cells, the number of metastasis and the survival days of mice were observed. Results: yCD3 reacted specifically with T cells, and 5μgyCD3 competitively inhibited the binding of 70% of standard anti-CD3 antibodies to cell-surface CD3 molecules. yCD3 stimulation of peripheral blood lymphocyte proliferation effective concentration of 8μg / L, and with IL-2, anti-CD28 antibody synergistic effect. The activated CD3AK cells in CD3 +, CD8 + and CD25 + cells increased; produce IL-2 and IFN γ CD3 + cells have varying degrees of increase in anti-CD28 antibody synergistic stimulation were increased by 3.29 and 2.47 times. 54%. When the effective target cell ratio of 80: 1, CD3AK cells on the percentage of tumor cell killing in vitro was 57.54%. 70%, inhibitory rate of tumor lung metastasis in mice was 39. 70%, with LAK cells combined treatment of more significant effect. Conclusion: yCD3 can activate T cells. The induced CD3AK cells show the effect of inhibiting tumor in vitro and in vivo, which is of great significance in the clinical anti-tumor adoptive immunotherapy.