Viale M, Ferrini S, Serrano S, Serrano D, Ardizzoni A, Nicolin A
Istituto Nazionale per la Ricerca sul Cancro, Laboratorio di Farmacologia, Genova, Italy.
Tumori. 1990 Oct 31;76(5):488-94. doi: 10.1177/030089169007600515.
Tumor infiltrating (TIL) and peripheral blood lymphocytes (PBL) were isolated from 18 patients with non-small cell lung cancer undergoing radical surgery. Surface marker analysis revealed that TILs and PBLs mainly consisted of CD3+ T cells and that TILs generally displayed a lower CD4/CD8 ratio. Differences were found in the expression of CD25 (IL-2 receptor) and DR (MHC class II) antigens, which were increased in TILs, and in the percentage of CD16+ natural killer (NK) cells, which was reduced in TILs as compared to PBLs. Accordingly, the NK activity of TILs was lower than that of PBLs, whereas neither TILs nor PBLs expressed spontaneous cytolytic activity against fresh autologous tumor cells, melanoma cells and the "NK-resistant" A549 lung carcinoma cell line. After 4 days of culture in medium with recombinant-interleukin-2 (rIL-2), TILs and PBLs acquired cytolytic activity against all cell targets, but TILs expressed higher levels of cytotoxicity than autologous PBLs only in 3 patients out of 16 tested. More importantly, both TILs and PBLs displayed similar levels of cytotoxic activity against autologous tumor cells. TILs and PBLs from 8 patients were also analyzed by a limiting dilution microculture system. Cloning efficiency was remarkably lower in TILs, and surface marker analysis of T cell clones confirmed that an accumulation of CD8+ lymphocytes, which displayed cytolytic activity in a lectin-dependent assay, occurred at the tumor site. The non-MHC-restricted cytolytic activity of TIL- and PBL-derived T cell clones against K562, A549, and allogeneic melanoma cells and the cytolytic activity against autologous tumor cells showed no significant differences. Only 53% of TIL clones released IL-2 in response to PHA + TPA stimulation, whereas 68% of PBL-derived clones were IL-2 producers. Moreover, most PBL- and TIL-derived clones released tumor necrosis factor alpha in response to mitogen stimulation.
从18例接受根治性手术的非小细胞肺癌患者中分离出肿瘤浸润淋巴细胞（TIL）和外周血淋巴细胞（PBL）。表面标志物分析显示，TIL和PBL主要由CD3 + T细胞组成，且TIL的CD4/CD8比值通常较低。在CD25（IL - 2受体）和DR（MHC II类）抗原的表达上发现了差异，这些抗原在TIL中增加，而CD16 +自然杀伤（NK）细胞的百分比在TIL中比PBL中降低。因此，TIL的NK活性低于PBL，而TIL和PBL均未表现出对新鲜自体肿瘤细胞、黑色素瘤细胞和“NK抗性”A549肺癌细胞系的自发细胞溶解活性。在含有重组白细胞介素-2（rIL - 2）的培养基中培养4天后，TIL和PBL获得了对所有细胞靶标的细胞溶解活性，但在16例测试患者中，只有3例TIL表现出比自体PBL更高水平的细胞毒性。更重要的是，TIL和PBL对自体肿瘤细胞表现出相似水平的细胞毒性活性。还通过有限稀释微量培养系统分析了8例患者的TIL和PBL。TIL的克隆效率明显较低，T细胞克隆的表面标志物分析证实，在肿瘤部位出现了CD8 +淋巴细胞的积累，这些淋巴细胞在凝集素依赖性试验中表现出细胞溶解活性。TIL和PBL来源的T细胞克隆对K562、A549和同种异体黑色素瘤细胞的非MHC限制性细胞溶解活性以及对自体肿瘤细胞的细胞溶解活性均无显著差异。只有53%的TIL克隆在PHA + TPA刺激下释放IL - 2，而68%的PBL来源克隆是IL - 2产生者。此外，大多数PBL和TIL来源的克隆在有丝分裂原刺激下释放肿瘤坏死因子α。
