Good manufacturing practices production of natural killer cells for immunotherapy: a six-year single-institution experience
Good manufacturing practices production of natural killer cells for immunotherapy: a six-year single-institution experience
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DOI：
10.1111/j.1537-2995.2006.01145.x
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发表时间：
2007-03-01
期刊：
影响因子：
2.9
通讯作者：
Miller, J. S.
中科院分区：
文献类型：
--
作者：
McKenna, D. H.;Sumstad, D.;Miller, J. S.
背景技术背景：自然杀伤（NK）细胞是淋巴细胞的一个子集，也是先天免疫系统的一部分，在防御癌症和病毒感染方面发挥着至关重要的作用。本文是一份关于临床规模的NK细胞治疗晚期癌症的良好生产规范（GMP）生产经验的报告。研究设计和方法：用细胞选择系统对未动员的外周血单个核细胞进行两种类型的NK细胞富集（CliniMACS，Miltenyi）：去除CD 3细胞以富集NK细胞，去除CD 3细胞后选择CD 56细胞以获得更纯的NK细胞产物。与白细胞介素-2（IL-2）过夜孵育后，细胞被洗涤，重新悬浮在5%的人血清白蛋白，然后释放infusion.RESULTS：自2000年以来，共生产了70种NK细胞治疗产品用于患者输液。对于去除CD 3细胞的NK细胞产物，平均纯度、回收率和活力分别为38%、79%和86%。对于去除CD 3细胞/富集CD 56细胞的NK细胞产物，平均纯度、回收率和活力分别为90%、19%和85%。革兰氏染色、无菌性和内毒素检测均在既定批放行的可接受限度内。与静息处理的细胞相比，IL-2活化显著增加了细胞在细胞毒性检测中的功能。结论：临床规模的NK细胞生产是有效的，可以在GMP下进行。纯化的NK细胞产物导致高NK细胞纯度，T细胞、单核细胞和B细胞的污染最小，但与单独去除CD 3细胞的NK细胞富集相比，其需要更多的处理时间，并导致较低的NK细胞回收率。额外的实验室研究和临床试验结果将确定NK细胞产品的最佳来源和类型。
BACKGROUND: Natural killer (NK) cells, a subset of lymphocytes and part of the innate immune system, play a crucial role in defense against cancer and viral infection. Herein is a report on the experience of clinical-scale, good manufacturing practices (GMPs) production of NK cells to treat advanced cancer.STUDY DESIGN AND METHODS: Two types of NK cell enrichments were performed on nonmobilized peripheral blood mononuclear cell apheresis collections with a cell selection system (CliniMACS, Miltenyi): CD3 cell depletion to enrich for NK cells and CD3 cell depletion followed by CD56 cell selection to obtain a more pure NK cell product. After overnight incubation with interleukin-2 (IL-2), cells were washed, resuspended in 5 percent human serum albumin, and then released for infusion.RESULTS: A total of 70 NK cell therapy products have been manufactured for patient infusion since 2000. For the CD3 cell-depleted NK cell products, the mean purity, recovery, and viability were 38, 79, and 86 percent, respectively. For the CD3 cell-depleted/CD56 cell-enriched NK cell products, the mean purity, recovery, and viability were 90, 19, and 85 percent, respectively. Gram stain, sterility, and endotoxin testing were all within acceptable limits for established lot release. Compared to the resting processed cells, IL-2 activation significantly increased the function of cells in cytotoxicity assays.CONCLUSION: Clinical-scale production of NK cells is efficient and can be performed under GMPs. The purified NK cell product results in high NK cell purity with minimal contamination by T cells, monocytes, and B cells, but it requires more time for processing and results in a lower NK cell recovery when compared to NK cell enrichment with CD3 cell depletion alone. Additional laboratory studies and results from clinical trials will identify the best source and type of NK cell product.