Zheng Ying-juan, Zhang Jian, Wang Sen-ming, Zhang Ji-ren
Center of Oncology, Zhujiang Hospital, First Military Medical Universility, Guangzhou 510282, China.
Di Yi Jun Yi Da Xue Xue Bao. 2004 Apr;24(4):392-6.
To understand the relation between cytotoxic activity of immunologic effector cells and multidrug resistance of the tumor cells.
Continuous observation of the morphological changes and MTT colorimetry were employed to evaluate the cytotoxic activity of lymphokine-activated killer (LAK) cells and natural killer (NK) cells against multidrug-resistant (MDR) human oral carcinoma cell line-KBV200 (before and after reversal of MDR) and parental drug-sensitive cell line KB. The morphologic changes of LAK cells and the 3 target cell lines were observed continuously under inverted microscope 3 h after co-culture of LAK cells with one of three target cell lines respectively. The lysis rates of three tumor cell lines in response to co-culture with LAK or NK cells were determined using MTT colorimetry.
In comparison with the parental drug-sensitive cell line KB, both KBV200 and its reserved cell line by verapamil (KBVV) showed earlier adherence and greater number of cells lysed by LAK. In MTT colorimetry assay, the cytotoxicity of both LAK and NK cells against the 3 cell lines was associated with the effector-to-target (E/T) cell ratio; the lysis rates of KBV200 and reversed KBV200 cells by verapamil in response to LAK and NK cells were higher than that of KB cells (P<0.05), but KBV200 and KBVV did not significantly differ (P>0.05). At the same E/T ratio, LAK cells possessed stronger cytotoxicity than NK cells against all the tumor cell lines (P<0.05).
Immunologic effector cells possess strong cytotoxic activity against multidrug-resistant cell line KBV200. Modulation of MDR does not decrease the cytotoxic activity of the immunologic effector cells. The results of this study suggest that adoptive cell immunotherapy with immunologic effector cells may be of value in controlling the progress of MDR tumors.
了解免疫效应细胞的细胞毒活性与肿瘤细胞多药耐药性之间的关系。
采用连续形态学观察和MTT比色法,评估淋巴因子激活的杀伤细胞(LAK细胞)和自然杀伤细胞(NK细胞)对多药耐药的人舌癌细胞系——KBV200(多药耐药逆转前后)及亲本药物敏感细胞系KB的细胞毒活性。分别将LAK细胞与三种靶细胞系之一共培养3小时后,在倒置显微镜下连续观察LAK细胞及三种靶细胞系的形态变化。采用MTT比色法测定三种肿瘤细胞系与LAK或NK细胞共培养后的裂解率。
与亲本药物敏感细胞系KB相比,KBV200及其经维拉帕米逆转后的细胞系(KBVV)被LAK细胞裂解的细胞贴壁更早且数量更多。在MTT比色法检测中,LAK和NK细胞对三种细胞系的细胞毒性均与效应细胞与靶细胞(E/T)比例相关;LAK和NK细胞对KBV200及经维拉帕米逆转后的KBV200细胞的裂解率高于KB细胞(P<0.05),但KBV200与KBVV之间差异无统计学意义(P>0.05)。在相同E/T比例下,LAK细胞对所有肿瘤细胞系的细胞毒性均强于NK细胞(P<0.05)。
免疫效应细胞对多药耐药细胞系KBV200具有较强的细胞毒活性。多药耐药的调节并不降低免疫效应细胞的细胞毒活性。本研究结果提示,采用免疫效应细胞的过继性细胞免疫治疗可能对控制多药耐药肿瘤的进展具有价值。