Mirjačić Martinović Katarina M, Babović Nada Lj, Džodić Radan R, Jurišić Vladimir B, Tanić Nikola T, Konjević Gordana M
Departments of aExperimental Oncology bMedical Oncology cSurgical Oncology Clinic, Institute of Oncology and Radiology of Serbia dSchool of Medicine, University of Belgrade eDepartment of Neurobiology, Institute for Biological Research 'Siniša Stanković', University of Belgrade, Belgrade fFaculty of Medical Sciences, University of Kragujevac, Kragujevac, Serbia.
Melanoma Res. 2014 Aug;24(4):295-304. doi: 10.1097/CMR.0000000000000072.
Although natural killer (NK) cells play an important antitumor role, melanoma cells may affect their effector functions. In this study, we analyzed the expression of various receptors and effector molecules in NK cells and their subsets in metastatic melanoma (MM) patients compared with healthy controls (HCs). In HC and MM patients, we analyzed NK cell activity using a chromium release assay and the expression of CD107a degranulation marker, activating NKG2D, NKp46, DNAM-1, and inhibitory CD158a and CD158b receptors, IL-12R beta 1, IL-12R beta 2, intracellular interferon (IFN)-γ, perforin, and STAT-1 in CD3-CD56+ NK cells, and cytotoxic CD3-CD56 and immunoregulatory CD3-CD56 subsets by flow cytometry. MM patients compared with HC not only had significantly decreased NK cell activity, lower expression of CD107a, and impaired IFN-γ production but also had decreased expression of activating NKG2D, NKp46, and DNAM-1 receptors, which was followed by lower expression of perforin, STAT-1, and both IL-12R subunits in NK cells. In MM patients only, there was a positive correlation between NKG2D expression and degranulation capacity, as well as IFN-γ production in NK cells. Analysis of the expression of various parameters of NK cell effector functions between MM patients with different localization of distant metastases showed that patients in the unfavorable M1c subclass had decreased expression of NKG2D and NKp46 on NK cells compared with patients in the M1a+b group. Downregulated NKG2D, NKp46, and DNAM-1 receptors associated with impaired NK cell effector function are important biomarkers of advanced disease with a poor prognosis in melanoma patients.
尽管自然杀伤（NK）细胞发挥着重要的抗肿瘤作用，但黑色素瘤细胞可能会影响其效应功能。在本研究中，我们分析了转移性黑色素瘤（MM）患者与健康对照（HC）相比，NK细胞及其亚群中各种受体和效应分子的表达情况。在HC和MM患者中，我们使用铬释放试验分析NK细胞活性，并通过流式细胞术分析CD3-CD56+NK细胞中CD107a脱颗粒标志物、活化性NKG2D、NKp46、DNAM-1以及抑制性CD158a和CD158b受体、IL-12Rβ1、IL-12Rβ2、细胞内干扰素（IFN）-γ、穿孔素和STAT-1的表达，以及细胞毒性CD3-CD56和免疫调节性CD3-CD56亚群。与HC相比，MM患者不仅NK细胞活性显著降低、CD107a表达降低且IFN-γ产生受损，而且活化性NKG2D、NKp46和DNAM-1受体的表达也降低，随后NK细胞中穿孔素、STAT-1以及IL-12R两个亚基的表达也降低。仅在MM患者中，NKG2D表达与脱颗粒能力以及NK细胞中IFN-γ产生之间存在正相关。对不同远处转移部位的MM患者NK细胞效应功能的各种参数表达进行分析表明，与M1a+b组患者相比，处于不利M1c亚类的患者NK细胞上NKG2D和NKp46的表达降低。与NK细胞效应功能受损相关的NKG2D、NKp46和DNAM-1受体下调是黑色素瘤患者晚期疾病预后不良的重要生物标志物。
