Institute for Oncology and Radiology of Serbia, Pasterova 14, 11000 Belgrade, Serbia.
Biomed Pharmacother. 2010 Dec;64(10):663-71. doi: 10.1016/j.biopha.2010.06.013. Epub 2010 Jul 17.
As IL-2 and IFN-α modulate NK cell activity it was of interest to investigate the expression of newly defined NK cell receptors and augmented NK cell activity in healthy individuals after cytokine in vitro treatment. Peripheral blood lymphocytes (PBL) obtained from 31 healthy volunteers treated for 18 h with 200 IU/ml IL-2 and 250 IU/ml IFN-α were evaluated for NK cell cytotoxicity. Expression of NKG2D, CD161, CD158a, CD158b receptors was analyzed on CD3⁻CD16+ NK cells, cytotoxic CD16(bright) and regulatory CD16(dim) subsets by FACS flow. The found induced significant in vitro enhancement of NK cell activity by both cytokines is supported by specific cytokine induction in PBL of pSTAT1 and pSTAT5, determined by Western blotting, as well as induction of IRF-1 transcription. Both cytokines induce significant up-regulation of NKG2D expression while only IFN-α induced significant up-regulation of CD161, with no alteration in KIR expression by either cytokine on CD3⁻CD16+ NK cells. Investigated cytokines did not induce change in NK cell bright and dim subset distribution. Moreover, we find that, not only cytokine receptor induction on the CD3⁻CD16+ NK cells, but also simultaneous increase in their percentage and/or density on CD16(bright) and CD16(dim) subsets, represent good indicators of receptor cytokine-susceptibility. As the role of NK cells has been shown in the loss of tolerance, infection and cancer, the data obtained in this study may be of help in NK cell profiling, by giving referent values of cytokine-induced novel NK cell receptor expression either in evaluation of these diseases or in immunomonitoring during cytokine immunotherapy.
由于 IL-2 和 IFN-α 可调节 NK 细胞的活性，因此研究健康个体在细胞因子体外处理后 NK 细胞新定义受体的表达和增强的 NK 细胞活性很有意思。用 200IU/ml IL-2 和 250IU/ml IFN-α 体外处理 31 名健康志愿者的外周血淋巴细胞（PBL）18 小时后，评估 NK 细胞的细胞毒性。通过 FACS 流式分析 CD3⁻CD16+ NK 细胞上 NKG2D、CD161、CD158a、CD158b 受体的表达，分析细胞毒性 CD16(bright)和调节性 CD16(dim)亚群。Western blot 确定 PBL 中 pSTAT1 和 pSTAT5 的特异性细胞因子诱导表明，两种细胞因子均能显著增强 NK 细胞的体外活性，而 IRF-1 转录的诱导也支持这一结果。两种细胞因子均能显著上调 NKG2D 的表达，而只有 IFN-α 能显著上调 CD161 的表达，两种细胞因子均不能改变 CD3⁻CD16+ NK 细胞上 KIR 的表达。研究的细胞因子不会改变 NK 细胞亮和暗亚群的分布。此外，我们发现，不仅 CD3⁻CD16+ NK 细胞上细胞因子受体的诱导，而且其在 CD16(bright)和 CD16(dim)亚群上的百分比和/或密度的同时增加，是受体细胞因子敏感性的良好指标。由于 NK 细胞的作用已在耐受丧失、感染和癌症中得到证实，因此本研究获得的数据可能有助于 NK 细胞分析，为评估这些疾病或细胞因子免疫治疗期间的免疫监测提供细胞因子诱导的新型 NK 细胞受体表达的参考值。
