慢性乙型肝炎患者PBMC中HBV特异性IL-21表达及其对CD8^+T淋巴细胞功能的影响  

Expression of HBV-specific IL-21 and its effect on the function of CD8^+T cells from peripheral blood mononuclear cells in patients with chronic hepatitis B

作　　者：成利伟 孔歌 李莉[1] 潘修成[1] 傅涓涓[1] 李丽[1] 

机构地区：[1]徐州医学院附属医院感染科,221000

出　　处：《肝脏》2014年第4期246-249,253,共5页Chinese Hepatology

基　　金：徐州医学院院长专项人才基金(2011KJZ18)

摘　　要：目的探讨慢性乙型肝炎患者外周血单个核细胞(PBMC)中HBV特异性IL-21的表达及其对CD8+T淋巴细胞功能的调节作用。方法以HBVc18-27(10μg/mL)多肽刺激46例不同临床类型慢性乙型肝炎患者PBMC。ELISA检测上清液IL-21水平。在HBVc18-27(10μg/mL)多肽和IL-21(50 U/mL)或IL-2(100 ng/mL)培养条件下,应用MHC-肽五聚体结合流式细胞术检测PBMC中HBV特异性CTL细胞频数变化;免疫磁珠分离纯化免疫耐受期和免疫激活期慢性乙型肝炎患者PBMC中CD8+T淋巴细胞,以HBcAg(10μg/mL)预刺激CD8-PBMC5 h后,采用Transwell小室共培养技术,在阻断IL-21通路条件下,应用实时PCR检测CD8+T淋巴细胞IFN-γmRNA的表达。结果 HBV特异性IL-21在免疫控制期和免疫激活期慢性乙型肝炎患者PBMC中表达水平显著高于免疫耐受期患者,而在免疫控制期和免疫激活期两组之间无显著差异。与PBS对照组相比,IL-21组HBV特异性CTL细胞频数明显增高,差异有统计学意义(P<0.01),而与IL-2组相比差异无统计学意义。以IL-21抗体阻断IL-21通路后,CD8+T淋巴细胞IFN-γmRNA表达水平显著下降,差异有统计学意义。结论免疫控制期和免疫激活期慢性乙型肝炎患者PBMC均可生成一定水平的HBV特异性IL-21,并能增强外周CD8+T淋巴细胞的增殖和IFN-γ表达水平。Objective To investigate the expression of HBV-specific IL-21 and its effect on the function of CD8＋T cells from peripheral blood mononuclear cells （PBMCs）in patients with chronic hepatitis B.Methods PBMCs was collected from 46 chronic hepatitis B （CHB）patients of different clinical types and was stimulated by HBVc18-27 peptide （10μg/mL）. IL-21 from supernatant was detected by ELISA. Stimulated by HBVc18-27 peptide （10μg/mL）,or in combination with IL-2 （50U/mL）or IL-21 （100ng/mL）,frequency of HBV specific CTL in CHB patients was evaluated by MHC-peptide pentamer in combination with flow cytometry assay. Pre-stimulated for 5 hours with HBcAg （10μg/mL）,isolated CD8＋T cells from PBMCs in immune-tolerant CHB patients co-cultured in transwell with isolated CD8-PBMCs derived from immune-active CHB patients. Level of IFN-γmRNA in CD8＋T cells was measured by real-time PCR when using anti-IL-21 antibody blocked the IL-21 pathway or not.Results Compared to immune-tolerant CHB patients,levels of HBV-associated IL-21 was much higher in immune-active and immune-inactive CHB patients. There was no significant difference of HBV---associated IL-21 levels between immune-active and immune-inactive CHB patients. Compared to PBS control group, frequency of HBV specific CTL from CHB patients was significantly higher in IL-21 group （P〈0.05）;however,there was no significant difference of frequency of HBV specific CTL between IL-21 group and IL-2 group. Level of IFN-γmRNA in CD8^＋ T cells was significantly decreased when using anti-IL-21 antibody blocked the IL-21 pathway.Conclusions IL-21 , which was much higher in immune-active and immune-inactive CHB patients,could enhance the HBV- specific CD8^＋ T cell＆amp;nbsp;function in patients with chronic hepatitis B.

关 键 词：病毒性肝炎 乙型 IL-21 CD8^+T 巴细胞 

分 类 号：R512.62[医药卫生—内科学]