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重组白细胞介素-15增强人脐带血自然杀伤细胞的细胞毒性。

Cytotoxicity of human cord blood natural killer cells is enhanced by recombinant interleukin-15.

作者信息

Saghafi Shiva, Pourfathollah Ali Akbar, Kheirandish Maryam, Azimdoust Anahita, Behnia Massoud, Shahjahani Mohammad, Moin Mostafa

机构信息

Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

出版信息

Iran J Allergy Asthma Immunol. 2010 Jun;9(2):69-77.

Abstract

Hematopoietic cord blood (CB) stem cell transplantation has more advantages to other cell sources because of lower Graft Versus Host Disease (GVHD). Interleukin-15(IL-15) is an immunoregulatory cytokine, known to enhance cytolytic function of cord Natural Killer (NK) cells. The aim of this study was to investigate the effect of IL-15 on NK cytotoxicity simultaneously in different cell death stages. We compared the ability of IL-15 to enhance the NK cytotoxicity of CB in comparison to adult blood Mononuclear Cells (MNCs) against K562 target cells by co-staining with AnnexinV-FITC and Propidium Iodide after 3.5 h incubation at 37 °C and 5% CO₂ by using flow cytometric method. We also evaluated phenotypic changes after treatment by IL-15 in both cell sources. Our results indicated that CB samples had lower level of apoptosis, while necrosis was negligible; also by escalating Effector: Target (E: T), we got higher level of apoptosis and necrosis in peripheral blood (PB). NK activity of cord and adult MNCs was enhanced by incubation with IL-15 (10 ng/ml) for 72 h with significantly higher results of PB in comparison to CB (p<0.0001). Moreover, IL-15 increased the percentage of CD3-/CD56+ and CD25+ cells after 72 h incubation. Results showed incubation with human recombinant (hr) IL-15 for 3 days increased NK activity. Taken together, these results indicated that NK cytotoxicity of CB MNCs could be augmented by human recombinant (hr) IL-15, but this activity did not reach to same level of PB counterparts. We established that CD25 expression on CB MNCs could be increased with IL-15, in 72-hour cultures, but to a lesser degree compared to that on corresponding adult PB MNCs.

摘要

造血脐带血(CB)干细胞移植相较于其他细胞来源具有更多优势,因为其移植物抗宿主病(GVHD)发生率较低。白细胞介素-15(IL-15)是一种免疫调节细胞因子,已知其可增强脐带自然杀伤(NK)细胞的溶细胞功能。本研究的目的是在不同细胞死亡阶段同时研究IL-15对NK细胞毒性的影响。我们通过流式细胞术,在37°C和5%二氧化碳条件下孵育3.5小时后,用膜联蛋白V-异硫氰酸荧光素(AnnexinV-FITC)和碘化丙啶共染色,比较了IL-15增强CB的NK细胞毒性与成人血单个核细胞(MNCs)针对K562靶细胞的能力。我们还评估了IL-15处理后两种细胞来源的表型变化。我们的结果表明,CB样本的凋亡水平较低,而坏死可忽略不计;此外,通过提高效应细胞:靶细胞(E:T)比例,我们在外周血(PB)中获得了更高水平的凋亡和坏死。脐带和成人MNCs与IL-15(10 ng/ml)孵育72小时后,NK活性增强,与CB相比,PB的结果显著更高(p<0.0001)。此外,孵育72小时后,IL-15增加了CD3-/CD56+和CD25+细胞的百分比。结果显示,与人重组(hr)IL-15孵育3天可增加NK活性。综上所述,这些结果表明,人重组(hr)IL-15可增强CB MNCs的NK细胞毒性,但这种活性未达到与PB对应细胞相同的水平。我们确定,在72小时培养中,IL-15可增加CB MNCs上CD25的表达,但与相应成人PB MNCs相比程度较小。

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