Saghafi Shiva, Pourfathollah Ali Akbar, Kheirandish Maryam, Azimdoust Anahita, Behnia Massoud, Shahjahani Mohammad, Moin Mostafa
Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Iran J Allergy Asthma Immunol. 2010 Jun;9(2):69-77.
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.
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。