Adler A, Albo V, Blatt J, Whiteside T L, Herberman R B
Pittsburgh Cancer Institute, University of Pittsburgh, PA.
Blood. 1989 Oct;74(5):1690-7.
Activation and expansion in culture with rIL-2 of peripheral blood (PB) and/or bone marrow (BM) specimens derived from children with ALL and ANLL, with active disease (AP) and in remission were studied (RP). Baseline NK cytolytic activity from AP was found to be depressed, whereas RP-derived cells had normal NK activity, as assayed against K562 targets. Culture in rIL-2 significantly enhanced the NK activity of both AP- and RP-derived cells and generated LAK activity, as assayed by 4-hour 51Cr release, against NK-resistant Raji cell line and against fresh, allogeneic, and autologous tumor cells. Lytic activity against fresh, cryopreserved leukemia blasts was of lower than that found against cell lines. In three patients higher lytic activity against autologous than against allogeneic blasts was demonstrated. Expansion in culture with rIL-2 varied from twofold to 120-fold. rIL-2 activation and expansion was better in RP than in AP. The predominant phenotype of activated cells, as determined by flow cytometry, was [mean % (SD)]: CD3- = 54 (12), CD8+ = 55 (17), and NKH1+ = 26 (7). The consistently high level of CD8+ cells was accompanied by very low levels of CD4+ cells: mean = 11% (14). Double-marker analysis showed mean of 33% (10) for CD3+/NKH1+ cells and mean = 32 (11) for CD8+/NKH1+ cells, implying that these populations were overlapping. Kinetics of expression of cell surface markers during 2 to 3 weeks in culture showed that CD8+ and NKH1+ enrichment occurred during the first week and lasted for up to 4 weeks, whereas CD4+ expression decreased after the second week. A significant decrease in the expression of IL-2 receptors (CD25) was observed from the second week of culture. This study shows the feasibility of in vitro generation of killer cells from PB and BM of pediatric leukemia patients.
研究了用重组白细胞介素-2（rIL-2）在体外培养来自急性淋巴细胞白血病（ALL）和急性非淋巴细胞白血病（ANLL）患儿的外周血（PB）和/或骨髓（BM）标本，这些患儿处于疾病活动期（AP）和缓解期（RP）。结果发现，与K562靶细胞相比，疾病活动期患儿的基线自然杀伤细胞（NK）溶细胞活性降低，而缓解期患儿来源的细胞具有正常的NK活性。用rIL-2培养可显著增强疾病活动期和缓解期患儿来源细胞的NK活性，并产生淋巴因子激活的杀伤细胞（LAK）活性，这通过4小时的51Cr释放试验来检测，该试验针对NK抗性的Raji细胞系以及新鲜的、同种异体和自体肿瘤细胞。对新鲜的、冻存的白血病原始细胞的溶细胞活性低于对细胞系的溶细胞活性。在3例患者中，显示出对自体原始细胞的溶细胞活性高于对同种异体原始细胞的溶细胞活性。用rIL-2在体外培养细胞的扩增倍数从2倍到120倍不等。rIL-2对缓解期患儿细胞的激活和扩增效果比对疾病活动期患儿细胞更好。通过流式细胞术确定，激活细胞的主要表型为[平均百分比（标准差）]：CD3- = 54（12），CD8+ = 55（17），NKH1+ = 26（7）。CD8+细胞持续高水平伴随着CD4+细胞的极低水平：平均值 = 11%（14）。双标记分析显示CD3+/NKH1+细胞的平均值为33%（10），CD8+/NKH1+细胞的平均值为32（11），这意味着这些群体存在重叠。在培养2至3周期间细胞表面标志物表达的动力学表明，CD8+和NKH1+的富集在第一周出现并持续长达4周，而CD4+表达在第二周后下降。从培养的第二周开始观察到白细胞介素-2受体（CD25）表达显著下降。本研究表明从儿童白血病患者的外周血和骨髓体外生成杀伤细胞是可行的。
