复发性生殖器疱疹患者外周血树突细胞的研究

Study on Peripheral Blood Dendritic Cells in Patients with Recurrent Genital Herpes

目的研究外周血树突细胞(DC)对疱疹病毒感染机体的反应和体外诱导成熟DC表面分子的表达及其意义。方法采用免疫荧光三标记流式细胞仪检测20例复发性生殖器疱疹(RGH)患者外周血DC亚群(CD11c+DC和CD123+DC)及用重组人粒细胞-巨噬细胞集落刺激因子、白介素4、肿瘤坏死因子α体外联合培养体系诱导成熟的DC表型(CD11c、CD123、CD1a、CD80、CD86、CD40和CD83等)的表达,并以18例正常人作为对照。结果RGH患者外周血CD11c+DC数量达2.54%±0.19%,比对照组(1.77%±0.83%)明显增高(P<0.05),CD123+DC(0.27%±0.16%)较对照组(0.43%±0.19%)显著降低(P<0.02);患者组CD1a、辅助刺激分子CD80及DC成熟的标志CD83等表达水平分别为0.21%±0.19%、0.28%±0.14%及0.18%±0.11%,显著高于对照组分别为0.06%±0.03%、0.17%±0.11%及0.10%±0.06%,P值分别为<0.01、<0.02及<0.01。RGH患者经体外细胞因子诱导成熟的DC表面各种免疫分子CD11c、CD123、CD1a、CD80、CD86、CD40和CD83的表达水平与对照组间差异均无显著性(P>0.05)。结论RGH患者外周血CD11c+DC摄取病毒抗原后被激活,表达高水平的辅助刺激分子,增强抗原提呈功能,直接启动抗病毒T细胞免疫应答。

Objective To investigate peripheral blood dendritic cells (DC) responding to herpesvirus infection in the patients with recurrent genital herpes and the expression of immune molecules on mature DC induced by the cytokines in vitro. Methods Using three-color flow cytometry, we studied the phenotype expression of CD11c, CD123, CDla, CD80, CD86, CD40 and CD83 on the two subsets of DC(CD11c+DC and CD123+DC) in peripheral blood. The monocytes isolated from the blood were cultured with recombinant human granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin-4 (IL-4) and tumor necrosis factor alpha (TNF-α) from 20 patients with recurrent genital herpes (RGH) and 18 healthy controls. Results The number of CD11c+ DC in the peripheral blood increased significantly in the peripheral blood cells of RGH patients (2.54% ± 1.19%) in comparison with that of healthy controls (1.77% ± 0.83%) (P < 0.05); the number of CD123+DC decreased in the RGH patients (0.27% ± 0.16%) was significantly lower than that in controls (0.43% ± 0.19%) (P < 0.02). The expression of CDla, co-stimulatory molecule(CD80) and the mature marker for DC (CD83), in the patients were 0.21% ± 0.19%, 0.28% ± 0.14% and 0.18% ± 0.11%, respectively, which were markedly higher than those in normal controls (0.06% ± 0.03%, 0.17% ± 0.11% and 0.10% ± 0.06%, respectively, P < 0.01, P < 0.02 and P < 0.01, respectively). In mature DC treated with cytokines, there was no significant difference in the levels of expression of various immune molecules (eg. CD11c, CD123, CD1a, CD80, CD86, CD40 and CD83 on mature DC) from the patients when compared with those in the controls (P > 0.05). Conclusions Peripheral blood CD11c+DC activated by uptaking viral antigens in the patients with RGH results in a high level expression of co-stimulatory molecules on DC and enhance antigen-presenting functions, which is essential for intiating antiviral immune response directly by T lymphocytes.