Directed Differentiation of Mobilized Hematopoietic Stem and Progenitor Cells into Functional NK Cells with Enhanced Antitumor Activity
Directed Differentiation of Mobilized Hematopoietic Stem and Progenitor Cells into Functional NK Cells with Enhanced Antitumor Activity
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DOI：
10.3390/cells9040811
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发表时间：
2020-04-01
期刊：
影响因子：
6
通讯作者：
Wels, Winfried S.
中科院分区：
文献类型：
--
作者：
Oberoi, Pranav;Kamenjarin, Kathrina;Wels, Winfried S.
获得足够数量的功能性自然杀伤（NK）细胞对于NK细胞过继免疫疗法的成功至关重要。虽然外周血（PB）扩增是目前选择的方法，但从造血干细胞和祖细胞（hsc）中体外生成NK细胞可能是一种有吸引力的替代方法。因此，动员到外周血中的造血干细胞（PB-CD34(+)）是一种有价值的起始材料，但早期研究中观察到的分离的PB-CD34（+）细胞向NK细胞的分化相当差且依赖供体，这仍然是一个主要障碍。在这里，我们报告了一种基于PB-CD34（+）细胞体外培养的改进方法，通过分析NK细胞相关的表面标记物和细胞毒性来评估，优化的细胞因子混合物可靠地产生功能成熟的NK细胞。为了进一步增强NK细胞的扩增，我们制备了K562饲养细胞，共表达4-1BB配体和膜锚定的IL-15和IL-21。pb来源的NK细胞和从造血干细胞离体分化的NK细胞与这些饲养细胞共同培养，显著提高了NK细胞的扩增，并完全补偿了仅基于细胞因子的繁殖过程中观察到的供体间差异。我们的研究结果表明，动员的PB-CD34（+）细胞根据这两步方案扩增和分化，作为产生异体NK细胞用于过继性癌症免疫治疗的有希望的来源。
Obtaining sufficient numbers of functional natural killer (NK) cells is crucial for the success of NK-cell-based adoptive immunotherapies. While expansion from peripheral blood (PB) is the current method of choice, ex vivo generation of NK cells from hematopoietic stem and progenitor cells (HSCs) may constitute an attractive alternative. Thereby, HSCs mobilized into peripheral blood (PB-CD34(+)) represent a valuable starting material, but the rather poor and donor-dependent differentiation of isolated PB-CD34(+) cells into NK cells observed in earlier studies still represents a major hurdle. Here, we report a refined approach based on ex vivo culture of PB-CD34(+) cells with optimized cytokine cocktails that reliably generates functionally mature NK cells, as assessed by analyzing NK-cell-associated surface markers and cytotoxicity. To further enhance NK cell expansion, we generated K562 feeder cells co-expressing 4-1BB ligand and membrane-anchored IL-15 and IL-21. Co-culture of PB-derived NK cells and NK cells that were ex-vivo-differentiated from HSCs with these feeder cells dramatically improved NK cell expansion, and fully compensated for donor-to-donor variability observed during only cytokine-based propagation. Our findings suggest mobilized PB-CD34(+) cells expanded and differentiated according to this two-step protocol as a promising source for the generation of allogeneic NK cells for adoptive cancer immunotherapy.