Ferrini S, Zarcone D, Viale M, Cerruti G, Millo R, Moretta A, Grossi C E
Istituto Nazionale per la Ricerca sul Cancro, Università di Genova, Italy.
Eur J Immunol. 1989 Jul;19(7):1183-8. doi: 10.1002/eji.1830190705.
The morphologic and functional characteristics of cells freshly isolated from human peripheral blood and bearing a T cell receptor (TcR) gamma/delta were analyzed. Cell preparations highly enriched for TcR gamma/delta+ cells were obtained by treatment of E rosette-forming lymphocytes with anti-CD4 and anti-CD8 monoclonal antibodies (mAb) and complement. These preparations consisted of 64-82% TcR gamma/delta+ lymphocytes, as indicated by the sum of cells reacting with the BB3 and A13 mAb which define two distinct, nonoverlapping, TcR gamma/delta+ cell subsets in the peripheral blood. TcR gamma/delta cells were able to form conjugates with the natural killer-sensitive K-562 and with the natural killer-resistant HL-60-R tumor cell lines. The cytochemical localization of lysosomal acid hydrolases showed that 95%-98% of the cells in the TcR gamma/delta+ preparations had the morphologic features of granular lymphocytes. Moreover, electron microscopy analyses showed that TcR gamma/delta+ cells had electron-dense granules dispersed in the cytoplasm and a variety of smooth vesicles, a morphology identical to that of other CD3- or CD3+ granular lymphocyte subsets. Freshly isolated TcR gamma/delta+ cells were unable to lyse K-562 and natural killer-resistant targets, such as HL-60-R and P815. However, low levels of target cell lysis were observed upon triggering of the effectors by anti-CD3 TcR mAb or by lectin. After short-term culture with interleukin 2, TcR gamma/delta+ cells acquired a strong cytolytic activity against K-562 and HL-60-R target cells in the absence of triggering stimuli, and also displayed high levels of cytolytic activity against P815 in the presence of anti-CD3/TcR mAb.
对从人外周血中新鲜分离出的带有T细胞受体(TcR)γ/δ的细胞的形态学和功能特性进行了分析。通过用抗CD4和抗CD8单克隆抗体(mAb)及补体处理E花环形成淋巴细胞,获得了高度富集TcRγ/δ+细胞的细胞制剂。这些制剂由64% - 82%的TcRγ/δ+淋巴细胞组成,这是通过与定义外周血中两个不同的、不重叠的TcRγ/δ+细胞亚群的BB3和A13 mAb反应的细胞总和来表明的。TcRγ/δ细胞能够与自然杀伤敏感的K - 562以及自然杀伤抗性的HL - 60 - R肿瘤细胞系形成结合物。溶酶体酸性水解酶的细胞化学定位显示,TcRγ/δ+制剂中95% - 98%的细胞具有颗粒淋巴细胞的形态特征。此外,电子显微镜分析表明,TcRγ/δ+细胞有电子致密颗粒分散在细胞质中以及各种光滑小泡,其形态与其他CD3 - 或CD3+颗粒淋巴细胞亚群相同。新鲜分离的TcRγ/δ+细胞无法裂解K - 562以及自然杀伤抗性靶细胞,如HL - 60 - R和P815。然而,在用抗CD3 TcR mAb或凝集素触发效应细胞后,观察到了低水平的靶细胞裂解。在用白细胞介素2进行短期培养后,TcRγ/δ+细胞在没有触发刺激的情况下获得了对K - 562和HL - 60 - R靶细胞的强大细胞溶解活性,并且在存在抗CD3/TcR mAb的情况下对P815也表现出高水平的细胞溶解活性。