Gelderblom Huub C, Nijhuis Laurens E J, de Jong Esther C, te Velde Anje A, Pajkrt Dasja, Reesink Henk W, Beld Marcel G H M, van Deventer Sander J H, Jansen Peter L M
Department of Gastroenterology and Hepatology, AMC Liver Center, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
Liver Int. 2007 Sep;27(7):944-53. doi: 10.1111/j.1478-3231.2007.01507.x.
Chronic hepatitis C virus (HCV) infection is characterized by an insufficient immune response, possibly owing to impaired function of antigen-presenting cells such as myeloid dendritic cells (DCs). Therapeutic vaccination with in vitro generated DCs may enhance the immune response. Subsets of DCs can originate from monocytes, but the presence of HCV in monocytes that develop into DCs in vitro may impair DC function. Therefore, we studied the presence of HCV RNA in monocytes and monocyte-derived DCs from chronic HCV patients.
Monocytes were cultured with granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin 4 (IL-4) for 6 days, and then with GM-CSF, IL-4, tumour necrosis factor-alpha (TNF-alpha), prostaglandin E2, IL-1beta and IL-6 for 2 days to generate mature DCs. HCV RNA was assessed by polymerase chain reaction. Surface molecules were assessed by flow cytometry. Cytokine production was assessed by cytokine bead array.
HCV RNA was present in monocytes in 11 of 13 patients, but undetectable in mature DCs in 13 of 13 patients. The morphology of patient DCs was comparable with DCs from healthy controls, but the percentage of cells expressing surface molecules CD83 (P=0.001), CD86 (P=0.023) and human leucocyte antigen-DR (P=0.028) was lower in HCV patients. Compared with control DCs, patient DCs produced enhanced levels of IL-10 (P=0.0079) and IL-8 (P=0.0079), and lower levels of TNF-alpha (P=0.032), IL-6 (P=NS) and IL-1beta (P=0.0079). Patient and control DCs did not produce IL-12.
Monocyte-derived DCs from chronic HCV patients are not infected but show an immature phenotype and aberrant cytokine profile.
慢性丙型肝炎病毒（HCV）感染的特征是免疫反应不足，这可能是由于抗原呈递细胞（如髓样树突状细胞（DCs））功能受损所致。用体外生成的DCs进行治疗性疫苗接种可能会增强免疫反应。DCs亚群可源自单核细胞，但体外发育成DCs的单核细胞中存在HCV可能会损害DC功能。因此，我们研究了慢性HCV患者单核细胞和单核细胞衍生DCs中HCV RNA的存在情况。
将单核细胞与粒细胞巨噬细胞集落刺激因子（GM-CSF）和白细胞介素4（IL-4）培养6天，然后与GM-CSF、IL-4、肿瘤坏死因子-α（TNF-α）、前列腺素E2、IL-1β和IL-6培养2天以生成成熟DCs。通过聚合酶链反应评估HCV RNA。通过流式细胞术评估表面分子。通过细胞因子微珠阵列评估细胞因子产生情况。
13例患者中有11例的单核细胞中存在HCV RNA，但13例患者的成熟DCs中均未检测到。患者DCs的形态与健康对照的DCs相当，但HCV患者中表达表面分子CD83（P = 0.001）、CD86（P = 0.023）和人类白细胞抗原-DR（P = 0.028）的细胞百分比更低。与对照DCs相比，患者DCs产生的IL-10（P = 0.0079）和IL-8（P = 0.0079）水平升高，而TNF-α（P = 0.032）、IL-6（P =无显著性差异）和IL-1β（P = 0.0079）水平降低。患者和对照DCs均不产生IL-12。
慢性HCV患者的单核细胞衍生DCs未被感染，但表现出不成熟的表型和异常的细胞因子谱。
