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目的探讨外周血浆样树突状细胞(pDC)Toll样受体(TLR)9的表达水平、pDC数量和分泌IFN-α能力的变化以及与慢性HBV感染状态的关系。方法流式细胞分析技术检测69例慢性HBV感染患者(包括HBV携带者14例,慢性乙型肝炎患者30例,乙型肝炎肝硬化患者25例)和21例健康对照组外周血pDC频率和TLR9表达水平;用CpG ODN 2216刺激外周血单个核细胞(PBMC),并与PBMC共培养24h,检测上清液中pDC产生IFN-α的能力;分析pDC数量、分泌IFN-α功能变化及其临床意义。应用SPSS 13.0进行统计学分析。结果与健康对照组(62.6±10.7)相比,HBV携带者、慢性乙型肝炎、乙型肝炎肝硬化患者pDC表达TLR9平均荧光强度均有不同程度下降(P<0.05)。与慢性乙型肝炎患者(54.3±16.9)比较.HBV携带者及乙型肝炎肝硬化患者pDC表达TLR9平均荧光强度分别为44.5±10.3和45.9±13.8,差异有统计学意义(P<0.05)。健康对照组外周血pDC细胞频率为(0.88±0.54)%,明显高于慢性乙型肝炎患者的(0.51±0.36)%和乙型肝炎肝硬化患者的(0.5±0.3)%(P<0.05)。慢性HBV感染患者外周血pDC频率与血清ALT水平呈明显负相关(r=-0.341.P<0.05)。HBV携带者、慢性乙型肝炎患者、乙型肝炎肝硬化患者PBMC中pDC分泌IFN-α的水平分别为(291.10±198.16)、(285.34±155.56)和(206.92±148.36)pg/mL,明显低于健康者的(600.23±344.91)pg/mL(P<0.05)。结论慢性HBV感染患者外周血pDC TLR9表达低下,外周血pDC频率和分泌IFN-α的能力明显降低,这可能是HBV持续感染的重要机制。
Objective To investigate the expression of TLR9, the number of pDC and the secretion of IFN-α in peripheral blood plasma-like dendritic cells (pDC) 9 and its relationship with chronic HBV infection. Methods Flow cytometry was used to detect the expression of pDC and TLR9 in 69 patients with chronic HBV infection (including 14 cases of HBV carriers, 30 cases of chronic hepatitis B, 25 cases of hepatitis B cirrhosis) and 21 healthy controls (PBMCs) were stimulated with CpG ODN 2216 and co-cultured with PBMC for 24 hours. The ability of pDC to produce IFN-α in the supernatant was assayed. The changes of pDC and the secretion of IFN-α and its clinical significance significance. SPSS 13.0 was used for statistical analysis. Results Compared with healthy controls (62.6 ± 10.7), the mean fluorescence intensity of pDR expressing TLR9 in HBV carriers, chronic hepatitis B and hepatitis B cirrhosis patients decreased to some extent (P <0.05). Compared with patients with chronic hepatitis B (54.3 ± 16.9), the average fluorescence intensity of TLR9 expression in pDC of patients with HBV and hepatitis B cirrhosis were 44.5 ± 10.3 and 45.9 ± 13.8, respectively (P <0.05). The frequency of pDC in the peripheral blood of healthy controls was (0.88 ± 0.54)%, significantly higher than that of patients with chronic hepatitis B (0.51 ± 0.36)% and (0.5 ± 0.3)% (P <0.05) . There was a negative correlation between pDC frequency and serum ALT level in patients with chronic HBV infection (r = -0.341, P <0.05). The levels of IFN-α secreted by pDC in PBMC of HBV carriers and chronic hepatitis B patients were (291.10 ± 198.16), (285.34 ± 155.56) and (206.92 ± 148.36) pg / mL, respectively, (600.23 ± 344.91) pg / mL in healthy subjects (P <0.05). Conclusion The expression of pDC TLR9 in peripheral blood of patients with chronic HBV infection is low, and the pDC frequency and the ability of secreting IFN-α in peripheral blood are obviously decreased, which may be an important mechanism of persistent HBV infection.