Zhou M X, Findley H W, Davis R, Ragab A H
Division of Pediatric Hematology/Oncology, Emory University, Atlanta, GA 30322.
Blood. 1990 Jan 1;75(1):160-5.
We recently reported that low molecular weight B-cell growth factor (LMW-BCGF) plus recombinant interleukin-2 (rIL-2) synergistically induced lymphokine-activated killer (LAK) activity from the bone marrow (BM) cells of children with acute lymphoblastic leukemia (ALL). The kinetics of cell growth, antigenic phenotype, and lytic activity of the generated effector cells were further analyzed in this study. BM cells from ALL patients with active disease and in complete remission (CR) were cultured with a combination of LMW-BCGF and rIL-2. Monoclonal antibodies (anti-CD3 and anti-Leu 19) and immunomagnetic beads were used to separate LAK cells into three subsets: CD3+/Leu 19-, CD3+/Leu 19+, and CD3-/Leu 19+. Cytotoxicity assays with different subsets were performed versus K562, Raji, and autologous leukemic cells, using a 3-hour 51Cr release test. There was a significant cell expansion of 54-fold (mean value) for CD3+ cells and 15-fold for Leu 19+ cells in culture with LMW-BCGF plus rIL-2 for 7 to 14 days, whereas no cell expansion was observed in culture with rIL-2 alone. Although NK activity (K562) was generated from leukemic BM cells in culture with rIL-2 alone, it is only about one third of that generated in culture with rIL-2 plus LMW-BCGF. Analysis of lytic activity of cells generated in the latter cultures demonstrated that CD3-/Leu 19+ cells expressed highest lytic activity against NK-sensitive K562 cells as well as against NK-resistant Raji cells. CD3+/Leu 19+ cells showed median cytotoxicity, and CD3+Leu 19- cells mediated only minimal cytotoxic activity. Also, lytic activity of CD3-/Leu 19+ cells against autologous leukemic blasts was noted in patients with active disease. Our results demonstrate that LAK activity generated from BM cells by LMW-BCGF and r-IL2 is mediated mainly by two types of Leu 19+ cells: CD3-/Leu 19+ NK cells and CD3-/Leu 19+ T cells. Although CD3+ T cells (both Leu 19+ and Leu 19-) mediated less antitumor cytotoxicity than CD3-/Leu 19+ cells, the former cells were the major expanding cell population in culture with LMW-BCGF and rIL-2. The new culture system may be effective in generation of cells with LAK activity for therapeutic use.
我们最近报道，低分子量B细胞生长因子（LMW - BCGF）加重组白细胞介素-2（rIL - 2）可协同诱导急性淋巴细胞白血病（ALL）患儿骨髓（BM）细胞产生淋巴因子激活的杀伤（LAK）活性。本研究进一步分析了所产生效应细胞的细胞生长动力学、抗原表型和裂解活性。将处于疾病活动期和完全缓解期（CR）的ALL患者的BM细胞与LMW - BCGF和rIL - 2组合进行培养。使用单克隆抗体（抗CD3和抗Leu 19）和免疫磁珠将LAK细胞分为三个亚群：CD3 + / Leu 19 -、CD3 + / Leu 19 +和CD3 - / Leu 19 +。使用3小时51Cr释放试验，对不同亚群针对K562、Raji和自体白血病细胞进行细胞毒性测定。在使用LMW - BCGF加rIL - 2培养7至14天的情况下，CD3 +细胞有显著的54倍（平均值）细胞扩增，Leu 19 +细胞有15倍扩增，而单独使用rIL - 2培养未观察到细胞扩增。虽然单独使用rIL - 2培养白血病BM细胞可产生NK活性（针对K562），但仅约为使用rIL - 2加LMW - BCGF培养所产生NK活性的三分之一。对后一种培养物中产生的细胞的裂解活性分析表明，CD3 - / Leu 19 +细胞对NK敏感的K562细胞以及对NK抗性的Raji细胞均表现出最高的裂解活性。CD3 + / Leu 19 +细胞表现出中等细胞毒性，而CD3 + Leu 19 -细胞仅介导最小的细胞毒性活性。此外，在疾病活动期患者中还观察到CD3 - / Leu 19 +细胞对自体白血病母细胞的裂解活性。我们的结果表明，LMW - BCGF和r - IL2从BM细胞产生的LAK活性主要由两种类型的Leu 19 +细胞介导：CD3 - / Leu 19 + NK细胞和CD3 - / Leu 19 + T细胞。虽然CD3 + T细胞（Leu + 19和Leu 19 -）介导的抗肿瘤细胞毒性低于CD3 - / Leu 19 +细胞，但前者细胞是使用LMW - BCGF和rIL - 2培养时主要的扩增细胞群体。这种新的培养系统可能有效地产生具有LAK活性的细胞用于治疗。
