Harwood Noah M K, Golden-Mason Lucy, Cheng Linling, Rosen Hugo R, Mengshol John A
Division of Gastroenterology and Hepatology, University of Colorado School of Medicine, Denver VA Medical Center, Denver, Colorado, USA.
Division of Gastroenterology and Hepatology, University of Colorado School of Medicine, Denver VA Medical Center, Denver, Colorado, USA
J Leukoc Biol. 2016 Mar;99(3):495-503. doi: 10.1189/jlb.5A1214-582R. Epub 2015 Oct 16.
The lectin galectin-9 may help establish and maintain chronic hepatitis C virus infection. Galectin-9 is elevated in the liver and sera of hepatitis C virus patients, induces apoptosis of hepatitis C virus-specific T cells, and increases inhibitory regulatory T cells. Kupffer cells stain strongly for galectin-9 protein in hepatitis C virus patients. In the current study, we determined stimuli that induce galectin-9 production by monocytes and macrophages in hepatitis C virus infection. With the use of real-time PCR and flow cytometry, we analyzed galectin-9 mRNA and protein from human monocytes cocultured with hepatitis C virus-infected cells or noninfectious hepatitis C virus subgenomic replicon cells. We focused on finding the stimuli for galectin-9 production. Additionally, we measured galectin-9 during monocyte-to-macrophage maturation. Finally, we examined galectin-9 in peripheral monocytes from hepatitis C virus patients using flow cytometry. Galectin-9 mRNA increased 8-fold when primary monocytes were exposed to hepatitis C virus--infected cells. Maximum induction required proximity or contact and did not require IFN-γ or hepatitis C virus virions. Coculture of monocytes with subgenomic replicon cells increased galectin-9 5-fold, and purified exosomes from infected cells stimulated galectin-9 production. Stimulation of monocyte TLR3, -7, and -8 increased galectin-9 production. Differentiation of monocytes to macrophages increased galectin-9, and nonclassic monocytes from hepatitis C virus patients had the highest levels of galectin-9. Hepatitis C virus-infected cells stimulated monocytes to produce galectin-9 in close proximity, possibly, in part, as a result of exosomes and endosomal TLRs. Differentiation of monocytes to macrophages increased galectin-9. Nonclassic monocytes from hepatitis C virus patients express the highest galectin-9 levels, suggesting they may contribute to elevated galectin-9 and adaptive immune inhibition in hepatitis C virus infection.
凝集素半乳糖凝集素-9可能有助于建立和维持丙型肝炎病毒感染。丙型肝炎病毒患者的肝脏和血清中半乳糖凝集素-9水平升高，可诱导丙型肝炎病毒特异性T细胞凋亡，并增加抑制性调节性T细胞。在丙型肝炎病毒患者中，库普弗细胞对半乳糖凝集素-9蛋白染色强烈。在本研究中，我们确定了丙型肝炎病毒感染中诱导单核细胞和巨噬细胞产生半乳糖凝集素-9的刺激因素。通过实时PCR和流式细胞术，我们分析了与丙型肝炎病毒感染细胞或非感染性丙型肝炎病毒亚基因组复制子细胞共培养的人单核细胞中的半乳糖凝集素-9 mRNA和蛋白。我们专注于寻找产生半乳糖凝集素-9的刺激因素。此外，我们在单核细胞向巨噬细胞成熟过程中测量了半乳糖凝集素-9。最后，我们使用流式细胞术检测了丙型肝炎病毒患者外周血单核细胞中的半乳糖凝集素-9。当原代单核细胞暴露于丙型肝炎病毒感染细胞时，半乳糖凝集素-9 mRNA增加了8倍。最大诱导需要接近或接触，且不需要干扰素-γ或丙型肝炎病毒颗粒。单核细胞与亚基因组复制子细胞共培养使半乳糖凝集素-9增加了5倍，从感染细胞中纯化的外泌体刺激了半乳糖凝集素-9的产生。刺激单核细胞的Toll样受体3、-7和-8可增加半乳糖凝集素-9的产生。单核细胞向巨噬细胞的分化增加了半乳糖凝集素-9，丙型肝炎病毒患者的非经典单核细胞中半乳糖凝集素-9水平最高。丙型肝炎病毒感染细胞在近距离刺激单核细胞产生半乳糖凝集素-9，这可能部分是由于外泌体和内体Toll样受体的作用。单核细胞向巨噬细胞的分化增加了半乳糖凝集素-9。丙型肝炎病毒患者的非经典单核细胞表达的半乳糖凝集素-9水平最高，这表明它们可能导致丙型肝炎病毒感染中半乳糖凝集素-9水平升高和适应性免疫抑制。
