Zhang Ting, Guo Chang-Jiang, Douglas Steven D, Metzger David S, O'Brien Charles P, Li Yuan, Wang Yan-Jian, Wang Xu, Ho Wen-Zhe
Department of Pediatrics, Division of Allergy and Immunology, Joseph Stokes Jr. Research Institute at The Children's Hospital of Philadelphia, PA 19104, USA.
Alcohol Clin Exp Res. 2005 Sep;29(9):1559-67. doi: 10.1097/01.alc.0000179364.32003.9f.
Natural killer (NK) cells are a critical component of the host innate immune system. We investigated whether alcohol impairs NK cell function, particularly production of CC chemokines induced by interleukin (IL)-2, the natural ligands for CCR5 receptor.
Primary NK cells and NK cell line (YTS) were cultured with or without alcohol (10 to 80 mM) for three hours. The culture supernatants were then harvested and used to treat human peripheral blood monocyte-derived macrophages and a HeLa cell line, which expresses CD4, CCR5, and CXCR4 receptors (MAGI cells). CC chemokine expression by YTS and primary NK cells treated with or without alcohol was analyzed with the real-time RT-PCR and ELISA. [Ca(2)(+)]i and Western blot assays were used to determine calcium-mediated intracellular signaling pathway and NF-kappaB p65 expression. HIV strains (Bal and UG024) were used to infect macrophages and MAGI cells. In addition, ADA (macrophage-tropic strain) and murine leukemia virus (MLV) envelope-pseudotyped HIV infection was carried out in macrophages. HIV infectivity was determined by HIV reverse transcriptase (RT) and beta-galactosidase activity assays.
Alcohol inhibited IL-2-induced CC chemokine (CCL3 and CCL4) expression by NK cells. Functional tests demonstrated that this reduced expression of CC chemokines was associated with diminished anti-HIV ability of NK cells. Alcohol also reduced the ability of NK cells to response to CCL3-mediated chemotaxis. Alcohol inhibited IL-2-induced NF-kappaB p65 protein expression and calcium mobilization by NK cells.
Alcohol, through the inhibition of IL-2-induced NF-kappaB p65 protein expression and intracellular calcium mobilization, suppressed NK cell production of CC chemokines. This suppression of CC chemokine production was associated with diminished anti-HIV activity of NK cells. Thus, by inhibiting NK cell-mediated innate immunity against HIV, alcohol consumption may have a cofactor role in the immunopathogenesis of HIV disease.
自然杀伤（NK）细胞是宿主固有免疫系统的关键组成部分。我们研究了酒精是否会损害NK细胞功能，尤其是白细胞介素（IL）-2诱导产生的CC趋化因子，CC趋化因子是CCR5受体的天然配体。
原代NK细胞和NK细胞系（YTS）在有或无酒精（10至80 mM）的条件下培养3小时。然后收集培养上清液，用于处理人外周血单核细胞衍生的巨噬细胞和表达CD4、CCR5和CXCR4受体的HeLa细胞系（MAGI细胞）。用实时逆转录聚合酶链反应（RT-PCR）和酶联免疫吸附测定（ELISA）分析有或无酒精处理的YTS和原代NK细胞中CC趋化因子的表达。采用钙离子浓度（[Ca(2)(+)]i）和蛋白质免疫印迹分析来确定钙介导的细胞内信号通路和核因子κB（NF-κB）p65的表达。使用HIV毒株（Bal和UG024）感染巨噬细胞和MAGI细胞。此外，在巨噬细胞中进行嗜巨噬细胞株ADA和鼠白血病病毒（MLV）包膜假型HIV感染。通过HIV逆转录酶（RT）和β-半乳糖苷酶活性测定来确定HIV感染性。
酒精抑制NK细胞IL-2诱导的CC趋化因子（CCL3和CCL4）表达。功能测试表明，CC趋化因子表达的降低与NK细胞抗HIV能力的减弱有关。酒精还降低了NK细胞对CCL3介导的趋化作用的反应能力。酒精抑制IL-2诱导的NK细胞NF-κB p65蛋白表达和钙动员。
酒精通过抑制IL-2诱导的NF-κB p65蛋白表达和细胞内钙动员，抑制NK细胞产生CC趋化因子。CC趋化因子产生受到的这种抑制与NK细胞抗HIV活性的减弱有关。因此，通过抑制NK细胞介导的针对HIV的固有免疫，饮酒可能在HIV疾病的免疫发病机制中起到辅助因子的作用。
