Fossati G, Anichini A, Squarcina P, Mazzocchi A, Parmiani G
Division of Experimental Oncology D, Istituto Nazionale Tumori, Milan, Italy.
Int J Cancer. 1988 Aug 15;42(2):239-45. doi: 10.1002/ijc.2910420216.
Peripheral blood lymphocytes (PBL) of a patient with metastatic melanoma were cultured with autologous melanoma cells (Auto-Me) and recombinant interleukin 2 (IL-2) (MLTC-PBL). Thirty-five days later, when no cytotoxicity against Auto-Me or K562 was detectable, MLTC-PBL were cloned in the presence of Auto-Me, IL-2 (25 U/ml) and Daudi cells as feeder. Eighty-one growing clones were simultaneously screened for proliferative and cytotoxic activity to Auto-Me. Twenty-two clones proliferated in the presence of Auto-Me only, 29 in the presence of IL-2 only and 41 in the presence of Auto-Me plus IL-2; 12 clones showed cytotoxic activity against Auto-Me. Six clones expressed both cytotoxic and proliferative activity to Auto-Me. The phenotype of 6 proliferative clones tested was CD3+, CD4+, WT31+, CD8-, CD16-, Leu19-, whereas that of 2 cytotoxic-proliferative clones tested was CD3+, CD8+, Leu19+, WT31+, CD4-, CD16-. Specificity analysis of proliferative response of 6 clones and of cytotoxicity of 7 clones, tested on a panel of 14 different target cells, revealed a complex pattern of reactivity, each clone expressing a peculiar specificity. Our results suggest the possibility of isolating, from melanoma patients' PBL, T-cell clones with proliferative activity to Auto-Me and Auto-Me plus IL-2, and T-cell clones which apparently express both proliferative and cytotoxic activity to Auto-Me.
将转移性黑色素瘤患者的外周血淋巴细胞(PBL)与自体黑色素瘤细胞(Auto-Me)和重组白细胞介素2(IL-2)一起培养(MLTC-PBL)。35天后,当检测不到对Auto-Me或K562的细胞毒性时,在Auto-Me、IL-2(25 U/ml)和作为饲养细胞的Daudi细胞存在的情况下对MLTC-PBL进行克隆。同时对81个生长的克隆进行针对Auto-Me的增殖和细胞毒性活性筛选。22个克隆仅在Auto-Me存在时增殖,29个仅在IL-2存在时增殖,41个在Auto-Me加IL-2存在时增殖;12个克隆对Auto-Me表现出细胞毒性活性。6个克隆对Auto-Me同时表现出细胞毒性和增殖活性。所测试的6个增殖克隆的表型为CD3+、CD4+、WT31+、CD8-、CD16- Leu19-,而所测试的2个细胞毒性增殖克隆的表型为CD3+、CD8+、Leu19+、WT31+、CD4-、CD16-。在一组14种不同靶细胞上对6个克隆的增殖反应和7个克隆的细胞毒性进行特异性分析,揭示了一种复杂的反应模式,每个克隆都表现出独特的特异性。我们的结果表明,有可能从黑色素瘤患者的PBL中分离出对Auto-Me和Auto-Me加IL-2具有增殖活性的T细胞克隆,以及对Auto-Me显然同时表现出增殖和细胞毒性活性的T细胞克隆。