Hutchinson G H, Heinemann D, Symes M O, Williamson R C
Br J Cancer. 1981 Sep;44(3):396-402. doi: 10.1038/bjc.1981.197.
Peripheral blood lymphocytes (PBL) were obtained from 13 patients and tumour-intrinsic lymphocytes (TIL) from 20 patients with colorectal cancer. The PBL were separated on a Ficoll-Isopaque gradient and the TIL by digestion of the tumour with collagenase-DNase. Both PBL and TIL were passed through nylon-wool columns and the eluted cells were co-cultured for 2 h with 51Cr-labelled tumour cells from the same patient. If patients in whom spontaneous 51Cr release from the tumour cells was greater than 33% were excluded, PBL showed cytotoxicity for the autoplastic tumour cells in 5/10 cases and TIL in 3/10 cases (NS). In 12 cases the cytotoxicity of the TIL was compared with that for TIL from the same tumour after the lymphocytes had been washed a further 6 times in Medium 199. Three effector: target (E/T) ratios, 5:1, 10:1 and 20:1, were used. The proportion of effector populations showing cytotoxicity was 2/12 for unwashed TIL and 9/12 for washed TIL (P less than 0.006). At the 5:1 E/T ratio the level of cytotoxicity was not significantly greater for washed TIL, but at the 10:1 ratio washed TIL showed significantly more cytotoxicity (P less than 0.025. At the 20:1 E/T ratio, a comparison was possible in 15 cases and the washed TIL again showed greater cytotoxicity (P less than 0.001).
从13例患者中获取外周血淋巴细胞(PBL),从20例结直肠癌患者中获取肿瘤固有淋巴细胞(TIL)。PBL通过Ficoll - Isopaque梯度分离,TIL通过用胶原酶 - 脱氧核糖核酸酶消化肿瘤获得。PBL和TIL均通过尼龙毛柱,洗脱的细胞与来自同一患者的51Cr标记肿瘤细胞共培养2小时。如果排除肿瘤细胞自发51Cr释放大于33%的患者,PBL在5/10例中对自身肿瘤细胞显示出细胞毒性,TIL在3/10例中显示出细胞毒性(无显著性差异)。在12例中,将淋巴细胞在199培养基中再洗涤6次后,比较TIL与来自同一肿瘤的TIL的细胞毒性。使用了三种效应细胞与靶细胞(E/T)比例,即5:1、10:1和20:1。未洗涤的TIL显示细胞毒性的效应细胞群体比例为2/12,洗涤后的TIL为9/12(P小于0.006)。在5:1的E/T比例下,洗涤后的TIL细胞毒性水平无显著升高,但在10:1比例下,洗涤后的TIL显示出显著更高的细胞毒性(P小于0.025)。在20:1的E/T比例下,15例中可以进行比较,洗涤后的TIL再次显示出更高的细胞毒性(P小于0.001)。