Natural-killer cell amplification for adoptive leukemia relapse immunotherapy: Comparison of three cytokines, IL-2, IL-15, or IL-7 and impact on NKG2D, KIR2DL1, and KIR2DL2 expression
Natural-killer cell amplification for adoptive leukemia relapse immunotherapy: Comparison of three cytokines, IL-2, IL-15, or IL-7 and impact on NKG2D, KIR2DL1, and KIR2DL2 expression
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DOI：
10.1016/j.exphem.2010.02.006
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发表时间：
2010-05-01
影响因子：
2.6
通讯作者：
Bensoussan, Daniele
中科院分区：
文献类型：
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作者：
Decot, Veronique;Voillard, Laure;Bensoussan, Daniele
Objective.自然杀伤（NK）细胞是一种淋巴细胞亚群，在造血干细胞移植环境中，介导移植物抗白血病效应，而无任何移植物抗宿主病。我们的目的是评估一种分离方法，可以使用良好的生产规范级试剂，并比较三种细胞因子的扩增，以设计未来的临床方案的基础上供体NK细胞输注治疗同种异体移植后复发。使用CD 3/CD 19耗竭方法富集NK细胞，并在2、10和50 ng/mL白细胞介素（IL）-2，IL-15或IL-7存在下扩增13天。在分离和培养后评价NK细胞的细胞毒性。在扩增过程中监测NKG 2D、KIR 2DL 2和KIR 2DL 1的表达。获得了高度T细胞和B细胞耗尽的NK细胞，并富集了2.6倍。用于扩增的最佳细胞因子浓度对于IL-2为10 ng/mL或对于IL-15为50 ng/mL。与10或50 ng/mL IL-2或与2、10或50 ng/mL IL-15孵育过夜后，以及与50 ng/mL IL-15孵育13天后，NK细胞的细胞毒性显著改善。使用IL-2和IL-15的组合没有显示出额外的益处，并且使用IL-7获得了阴性结果。培养后NK细胞的3种受体均明显上调，主要与IL-2或IL-15有关。在我们的研究中，10 ng/mL IL-2或50 ng/mL IL-15是扩增的最佳浓度，并且在显著增强细胞毒性和改变NK细胞受体表达模式方面是等同的。(C)2010 ISEH -血液学和干细胞学会。爱思唯尔公司出版
Objective. Natural killer (NK) cells are a lymphocyte subset that, in a hematopoietic stem cell transplantation setting, mediates a graft-vs-leukemia effect without any graft-vs-host disease. We aimed to evaluate an isolation method that can be used with Good Manufacturing Practices grade reagents and to compare three cytokines for expansion in order to design future clinical protocols based on donor NK-cell infusions to cure relapse after allograft.Materials and Methods. NK cells were enriched using a CD3/CD19 depletion method and expanded for 13 days in the presence of 2, 10, and 50 ng/mL interleukin (IL)-2, IL-15, or IL-7. NK-cell cytotoxicity was evaluated after isolation and culture. Expression of NKG2D, KIR2DL2, and KIR2DL1 was monitored during expansion.Results. Highly T- and B-cell depleted NK cells were obtained and enriched 2.6-fold. The optimal cytokine concentration for expansion was 10 ng/mL for IL-2 or 50 ng/mL for IL-15. NK-cell cytotoxicity was significantly improved after an overnight incubation with 10 or 50 ng/mL IL-2 or with 2, 10, or 50 ng/mL IL-15, and after 13 days with 50 ng/mL IL-15. The use of a combination of IL-2 and IL-15 showed no additional benefit and negative results were obtained with IL-7. The three NK cell receptors were significantly upregulated after culture, mainly with IL-2 or IL-15.Conclusion. In our study, 10 ng/mL IL-2 or 50 ng/mL, IL-15 were the optimal concentrations for expansion and were equivalent in significantly enhancing cytotoxicity and modifying NK-cell receptor expression patterns. (C) 2010 ISEH - Society for Hematology and Stem Cells. Published by Elsevier Inc.