Uchida A, Moore M
J Natl Cancer Inst. 1984 Dec;73(6):1285-92.
The specific and natural killer (NK)-restricted nature of autologous tumor killing by blood lymphocytes was studied in patients with carcinomatous pleural effusions. Large granular lymphocytes (LGL) and small T-lymphocytes were isolated by centrifugation on discontinuous Percoll density gradients. Tumor cells freshly isolated from pleural effusions of cancer patients were classified according to their susceptibility to purified LGL from normal donors in a 4-hour 51Cr release assay. Of 15 NK-sensitive tumors, 14 were lysed by fresh autologous LGL, whereas only 2 were killed by T-cells. Neither LGL nor T-cells were cytotoxic to NK-resistant autologous tumor. LGL and T-cells were then cultured in vitro with autologous tumor cells for 6 days. In 13 of 15 autologous mixed lymphocyte-tumor cultures (MLTC) NK-sensitive tumor-cultured LGL maintained their autotumor killing activity, whereas LGL cultured alone lost the activity. Depletion of high-affinity sheep erythrocyte-rosetting cells from Percoll-purified LGL resulted in an enrichment of effector cells. LGL from autologous MLTC were able to kill NK-susceptible allogeneic effusion tumor and K562 as were fresh LGL. No lysis of NK-resistant autologous tumor was observed with cultured LGL. In contrast, activation of T-cells in autologous MLTC resulted in the generation of autotumor killer cells in 10 of 15 NK-sensitive and 3 of 6 NK-resistant tumor samples. However, cultured T-cells were incapable of killing allogeneic tumor and K562. In autologous MLTC T-cells proliferated in response to autologous tumor, whereas no proliferation was observed in the culture of LGL. The enrichment of blasts from cultured T-cells on discontinuous Percoll gradients induced an augmentation of autotumor cytotoxicity, with no reactivity in blast-depleted, small, resting T-lymphocytes. These results indicated that 2 distinct types of autotumor-recognizing lymphocytes, LGL and T-cells, are present in the peripheral blood of cancer patients.
对癌性胸腔积液患者血液淋巴细胞对自体肿瘤的杀伤作用的特异性及自然杀伤（NK）细胞受限特性进行了研究。通过在不连续的Percoll密度梯度上离心分离出大颗粒淋巴细胞（LGL）和小T淋巴细胞。在4小时的51Cr释放试验中，根据癌症患者胸腔积液中新鲜分离的肿瘤细胞对正常供体纯化LGL的敏感性对其进行分类。在15个NK敏感肿瘤中，14个被新鲜的自体LGL裂解，而只有2个被T细胞杀伤。LGL和T细胞对NK抗性自体肿瘤均无细胞毒性。然后将LGL和T细胞与自体肿瘤细胞在体外培养6天。在15个自体混合淋巴细胞-肿瘤培养物（MLTC）中的13个中，NK敏感肿瘤培养的LGL保持其自体肿瘤杀伤活性，而单独培养的LGL则失去活性。从Percoll纯化的LGL中去除高亲和力绵羊红细胞花环形成细胞导致效应细胞富集。来自自体MLTC的LGL能够像新鲜LGL一样杀伤NK敏感的同种异体胸腔积液肿瘤和K562。培养的LGL未观察到对NK抗性自体肿瘤的裂解。相反，自体MLTC中T细胞的激活导致15个NK敏感肿瘤样本中的10个和6个NK抗性肿瘤样本中的3个产生自体肿瘤杀伤细胞。然而，培养的T细胞无法杀伤同种异体肿瘤和K562。在自体MLTC中，T细胞对自体肿瘤有增殖反应，而在LGL培养物中未观察到增殖。通过在不连续的Percoll梯度上培养T细胞使母细胞富集可增强自体肿瘤细胞毒性，而去除母细胞的小的静止T淋巴细胞则无反应性。这些结果表明，癌症患者外周血中存在两种不同类型的识别自体肿瘤的淋巴细胞，即LGL和T细胞。
