Kraaij Marina D, Vereyken Elly J F, Leenen Pieter J M, van den Bosch Thierry P P, Rezaee Farhad, Betjes Michiel G H, Baan Carla C, Rowshani Ajda T
Department of Nephrology and Transplantation, Erasmus University Medical Center, Rotterdam, The Netherlands.
Department of Immunology, Erasmus University Medical Center, Rotterdam, The Netherlands.
Cytokine. 2014 May;67(1):7-12. doi: 10.1016/j.cyto.2014.02.001. Epub 2014 Feb 26.
Representing a crucial T-helper 1 cytokine, IFN-γ acts as an important bridge between innate and adaptive immunity and is involved in many acute and chronic pathologic states, such as autoimmune diseases and solid organ transplant rejection. At present, debate still prevails about the ability of human monocytes to produce IFN-γ. We aimed to investigate whether human monocytes possess the capacity to produce IFN-γ at mRNA and protein level. Using real time PCR, flow cytometric analysis and ELISA, we investigated the capacity of freshly isolated CD14+ monocytes of healthy individuals and kidney transplant recipients to produce IFN-γ after stimulation with IFN-γ and LPS or LPS alone. We observed increased IFN-γ mRNA levels in CD14+ monocytes after stimulation as compared to the unstimulated controls in both populations. In addition, stimulation with IFN-γ and LPS or LPS alone led to a significant increase in the percentage of CD14+ monocytes producing TNF-α and IFN-γ at protein level (p<0.05). A trend towards increased secreted IFN-γ production in supernatants was also observed after LPS stimulation using ELISA. We conclude that human monocytes from healthy individuals and kidney transplant recipients possess the capacity to produce IFN-γ.
作为一种关键的辅助性T细胞1细胞因子，干扰素-γ（IFN-γ）是先天性免疫和适应性免疫之间的重要桥梁，并参与许多急性和慢性病理状态，如自身免疫性疾病和实体器官移植排斥反应。目前，关于人类单核细胞产生IFN-γ的能力仍存在争议。我们旨在研究人类单核细胞在mRNA和蛋白质水平上是否具有产生IFN-γ的能力。我们使用实时聚合酶链反应（PCR）、流式细胞术分析和酶联免疫吸附测定（ELISA），研究了健康个体和肾移植受者新鲜分离的CD14+单核细胞在受到IFN-γ和脂多糖（LPS）刺激或仅LPS刺激后产生IFN-γ的能力。我们观察到，与未刺激的对照组相比，刺激后两个群体中CD14+单核细胞的IFN-γ mRNA水平均有所升高。此外，用IFN-γ和LPS或仅用LPS刺激导致蛋白质水平上产生肿瘤坏死因子-α（TNF-α）和IFN-γ的CD14+单核细胞百分比显著增加（p<0.05）。使用ELISA法还观察到LPS刺激后上清液中分泌的IFN-γ产量有增加的趋势。我们得出结论，健康个体和肾移植受者的人类单核细胞具有产生IFN-γ的能力。
