Abstract

Preliminary results of clinical trials show that the use of activated and expanded ex vivo natural killer cells is a promising tool for adoptive immunotherapy of acute myeloid leukemia, as well as a number of solid tumors, when used together with targeted monoclonal antibodies. However, the introduction of this approach into clinical practice is limited by the possibility of obtaining a sufficient amount of the NK cell product with required characteristics. To solve this problem, we obtained transgenic feeder cells based on immortalized K562 cells expressing the recombinant membrane-bound variant of human interleukin-21 and 4-1BBL protein. Cocultivation of mononuclear cells obtained from the peripheral blood of ten healthy donors with genetically modified feeder cells resulted in significant expansion of natural killer cells (median expansion fold 21 589 times; minimum, 3150 times; maximum, 304 328 times) with a minimum content of T-cells (median, 0.5%; minimum, 0.06%; maximum, 4.7%) compared with their initial number. NK cells obtained in this way were not contaminated with the BCR–ABL1 oncogene from feeder cells and had a lower expression of c-MYC protooncogene compared to the initial level.

中文翻译：

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转基因饲养细胞存在下人类自然杀伤细胞的扩增和激活

摘要

临床试验的初步结果表明，与靶向单克隆抗体一起使用时，活化和扩增的离体自然杀伤细胞的使用对于急性髓细胞白血病以及许多实体瘤的过继免疫疗法是一种有前途的工具。然而，由于获得足够量的具有所需特性的NK细胞产物的可能性限制了将该方法引入临床实践。为了解决这个问题，我们基于永生化的K562细胞获得了转基因饲养细胞，该细胞表达了人白介素21和4-1BBL蛋白的重组膜结合变体。从十个健康供体的外周血中获得的单核细胞与转基因饲养细胞共培养，导致自然杀伤细胞显着扩增（中值扩增倍数为21 589倍；最小为3150倍；最大为304 328倍），其最小含量为与初始数量相比，T细胞（中位数为0.5％；最小值为0.06％；最大值为4.7％）。用这种方法获得的NK细胞没有被来自饲养细胞的BCR-ABL1癌基因污染，与初始水平相比，其c-MYC原癌基因的表达较低。