白细胞介素-15 在长期培养中支持生成高效的临床级自然杀伤细胞，用于靶向血液系统恶性肿瘤。

Interleukin-15 supports generation of highly potent clinical-grade natural killer cells in long-term cultures for targeting hematological malignancies.

机构信息

Blood Services Group, Health Sciences Authority, Singapore.

出版信息

Exp Hematol. 2011 Sep;39(9):904-14. doi: 10.1016/j.exphem.2011.06.003. Epub 2011 Jun 13.

DOI:10.1016/j.exphem.2011.06.003

PMID:21703984

Abstract

OBJECTIVE

Interleukin (IL)-15 is a promising novel cytokine for natural killer (NK) cell activation and survival. We studied the effects of IL-15 compared to IL-2 on NK cells in long-term cultures for clinical translation.

MATERIALS AND METHODS

CD56(+)CD3(-) NK cells were expanded with IL-2 or IL-15 for 2 to 4 weeks within lymphokine-activated killer (LAK) cell cultures (LAK-NK) in serum-enriched AIM V or CellGro Stem Cell Growth Medium (SCGM). Cell growth, viability, and NK cell content were monitored and cytotoxicity assessed in a flow cytometric cytotoxicity assay.

RESULTS

IL-15 (100-1000 U/mL) could replace IL-2 (1000 U/mL) in AIM V cultures to achieve efficient LAK cell expansion. However, IL-15-stimulated LAK cells exceeded cytotoxicity of IL-2-stimulated LAK cells against K562, notably at later culture points. In the powerful CellGro SCGM, LAK cells expanded over 28 days an average of 905-fold ± 320-fold standard error of the mean (SEM) for IL-2 (500 U/mL) and 484-fold ± 98-fold SEM for IL-15 (500 U/mL), and NK cells within such LAK cultures expanded an average of 2320-fold ± 975-fold SEM for IL-2 and 1084-fold ± 309-fold SEM for IL-15. Importantly, such IL-15-activated LAK-NK cells retained enhanced cytotoxicity at later culture points against K562 as well. IL-15-stimulated effectors were also highly cytotoxic against hematological targets MOLT-4 and KU812 and nontoxic against autologous nonmalignant cells. Interestingly, IL-15-LAK-NK cells showed overall significant upregulation of the main activating and inhibitory NK cell receptors after long-term cytokine stimulation.

CONCLUSIONS

Our results demonstrate the potential for IL-15 to support large-scale expansion of clinical-grade LAK-NK effectors, which could retain enhanced longer-term potency and preserve activation receptors in therapy of hematological malignancies. Protocols are readily clinically translatable.

摘要

目的

白细胞介素（IL）-15 是一种有前途的新型细胞因子，可用于自然杀伤（NK）细胞的激活和存活。我们研究了与 IL-2 相比，IL-15 对长期培养中的 NK 细胞的影响，以进行临床转化。

材料和方法

CD56(+)CD3(-)NK 细胞在富含血清的 AIM V 或 CellGro 干细胞生长培养基（SCGM）中进行淋巴因子激活的杀伤（LAK）细胞培养（LAK-NK），用 IL-2 或 IL-15 培养 2 至 4 周。监测细胞生长、活力和 NK 细胞含量，并通过流式细胞术细胞毒性测定评估细胞毒性。

结果

IL-15（100-1000 U/mL）可替代 AIM V 培养中的 IL-2（1000 U/mL），以实现高效的 LAK 细胞扩增。然而，与 IL-2 刺激的 LAK 细胞相比，IL-15 刺激的 LAK 细胞对 K562 的细胞毒性明显更高，尤其是在较晚的培养点。在强大的 CellGro SCGM 中，LAK 细胞在 28 天内平均扩增 905 倍±320 倍标准误差（SEM），对于 IL-2（500 U/mL）和 484 倍±98 倍 SEM 对于 IL-15（500 U/mL），并且此类 LAK 培养物中的 NK 细胞平均扩增 2320 倍±975 倍 SEM 对于 IL-2 和 1084 倍±309 倍 SEM 对于 IL-15。重要的是，这种 IL-15 激活的 LAK-NK 细胞在后期培养点也保留了对 K562 的增强细胞毒性。IL-15 刺激的效应物对血液学靶标 MOLT-4 和 KU812 也具有高度细胞毒性，而对自身非恶性细胞无毒。有趣的是，IL-15-LAK-NK 细胞在长期细胞因子刺激后表现出主要激活和抑制性 NK 细胞受体的总体显著上调。