Firouzi Javad, Hajifathali Abbas, Azimi Masoumeh, Parvini Neda, Ghaemi Fatemeh, Shayan Asl Niloufar, Hedayati Asl Amir Abbas, Safa Majid, Ebrahimi Marzieh
Department of Tissue Engineering and Regenerative Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran.
Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
Cell J. 2023 Feb 1;25(2):92-101. doi: 10.22074/cellj.2023.561054.1123.
Natural killer (NK) cells are critical immune cells for acute myeloid leukemia (AML) targeting. However, little is known about the relationship between using checkpoint inhibitors and heat shock protein 70 (Hsp70) as NK cell activators to control AML. Therefore, the study aims to find the best formulation of Hsp70, human PD-1 (Programmed cell death protein 1) blocker, and interleukin 15 (IL-15) to activate NK cells against AML.
In this experimental study, the NK cells were isolated from mononuclear cells (MNCs) by using magnetic activation cell sorting (MACS) and were activated using the different combinations of Hsp70, PD-1 blocker, and IL-15 and then followed by immunophenotyping, functional assays to estimate their killing potential, and evaluation of expression pattern of and A and B.
The expression of PD-1 was significantly (P<0.05) reduced after NK cell activation by the different formulas of IL-15, Hsp70, and PD-1 blocker. The expression of NKG2A in the treated NK cells was reduced particularly in the IL-15 (P<0.01) and IL-15+PD-1 blocker (P<0.05) groups. The addition of Hsp70 increased its expression. The cytotoxic effect of NK cells increased in all groups, especially in IL-15+PD-1 blocker besides increasing interferon-gamma (IFN-γ), Granzymes, and perforin expression (P<0.05). All IL-15+PD-1 blocker group changes were associated with the upregulation of and as key factors of NK cell activation. The presence of Hsp70 reduced IFN-γ releasing, and down-regulation of Granzymes, and Perforin (P<0.05).
We suggested the combination of IL-15 and PD-1 blocker could enhance the killing potential of AMLNK cells. Moreover, Hsp70 in combination with IL-15 and PD-1 blocker interferes activation of AML-NK cells through unknown mechanisms.
自然杀伤（NK）细胞是靶向急性髓系白血病（AML）的关键免疫细胞。然而，关于使用检查点抑制剂和热休克蛋白70（Hsp70）作为NK细胞激活剂来控制AML之间的关系，我们了解甚少。因此，本研究旨在寻找Hsp70、人程序性死亡蛋白1（PD - 1）阻断剂和白细胞介素15（IL - 15）激活NK细胞对抗AML的最佳配方。
在本实验研究中，通过磁激活细胞分选（MACS）从单核细胞（MNCs）中分离出NK细胞，并使用Hsp70、PD - 1阻断剂和IL - 15的不同组合对其进行激活，随后进行免疫表型分析、功能测定以评估其杀伤潜力，并评估 和 A和B的表达模式。
通过IL - 15、Hsp70和PD - 1阻断剂的不同配方激活NK细胞后，PD - 1的表达显著降低（P < 0.05）。在经处理的NK细胞中，NKG2A的表达降低，特别是在IL - 15组（P < 0.01）和IL - 15 + PD - 1阻断剂组（P < 0.05）。添加Hsp70可增加其表达。所有组中NK细胞的细胞毒性作用均增强，特别是在IL - 15 + PD - 1阻断剂组，同时干扰素 - γ（IFN - γ）、颗粒酶和穿孔素的表达也增加（P < 0.05）。IL - 15 + PD - 1阻断剂组的所有变化均与作为NK细胞激活关键因子的 和 的上调有关。Hsp70的存在降低了IFN - γ的释放，并下调了颗粒酶和穿孔素（P < 0.05）。
我们认为IL - 15和PD - 1阻断剂的组合可增强AML - NK细胞的杀伤潜力。此外，Hsp70与IL - 15和PD - 1阻断剂联合使用会通过未知机制干扰AML - NK细胞的激活。
