中文摘要：

目的研究各类肝硬化患者Th1／Th22失衡的特点及其与病程进展的关系，探索靶向治疗肝硬化的免疫治疗策略。方法体外离心法分离外周血单个核细胞，收集细胞进行CD3-BV500及CD8-PerCP—Cy5．5染色，再加入干扰素γ（IFNγ）-PE—Cy7、白细胞介素（IL）-17a—APC和IL-22-PE或者相应的同型对照，破膜洗液洗涤1次后加1％多聚甲醛固定。使用Flowjo分析软件进行T淋巴细胞亚群及细胞因子分析。定义Th1（CD4＋IFNγ＋），Th17（CD4＋IL-17a＋），Th22（CD4＋IL-22＋）以及Tc1（CD8＋IFNγ＋），Tc17（CD8＋IL-17a＋），Tc22（CD8＋IL-22＋），调查上述6个亚群分泌IFNγ、IL-17a、IL-22的情况。LX-2细胞无血清培养，添加不同浓度（25、50、100ng／ml）的重组人IL-22重组蛋白；24h后用活化指标α-平滑肌肌动蛋白检测LX-2活化情况，体积分数10％胎牛血清作为阳性对照。用酶联免疫吸附试验（化学发光法）检测上清液中透明质酸、III型胶原前体、IV型胶原浓度。据资料不同分别采用one-way ANOVA、Mann-Whitney非参数U检验或Kruskal wallisH非参数检验进行统计学分析。结果各类病因引起的肝硬化患者外周血Tc1、Th17和Th22细胞均较健康对照组显著增加。所有肝硬化患者Th17细胞频率（4．25％±2．45％）较对照组（2．59％±1．36％）增加了0．64倍，而肝硬化患者Th22细胞平均频率（4．17％±2．55％）较对照组（1．31％±0．64％）增加了2．18倍;尤其是酒精性肝硬化患者，其外周Th17（5．89％±3．44％）和Th22亚群（5．32％±3．67％）较对照组分别增加了1．27倍和3．06倍，P〈0．05。特别是酒精性肝硬化患者，其Th22细胞显著增加。抗纤维化和促纤维化因子（Th1／Th22）比值在各类肝硬化患者均显著降低，并与肝硬化严重程度呈正相关，是各类病因引起的肝硬化共同变化的免疫学特征。进一步证实，IL-22能激活肝星状?

英文摘要：

Objective To investigate the clinical features of imbalance between Th1 and Th22 cells and its association with disease progression in patients with liver cirrhosis, and to explore immune therapeutic strategies for targeted therapy for liver cirrhosis. Methods In vitro peripheral blood mononucleated cells （PBMCs） were collected by centrifugation. CD3-BV500 and CD8-PerCP-Cy5.5 staining was performed for these cells. IFNγ-PE-Cy7, IL-17a-APC, IL-22-PE, or the corresponding isotype control was added, and then PBMCs were fixed with 1% polyoxymethylene after being washed once by permeabilization-wash buffer.Flowjo softvare was used for the analysis of T lymphocyte subsets and cytokines. Th1 （CD4＋IFNγ＋）, Th17 （CD4＋IL-17a＋）, Th22 （CD4＋IL-22＋）, Tcl （CD8＋IFNγ＋）, Tcl7 （CD8＋IL-17a＋）, and Tc22 （CD8＋IL-22＋） subsets were defined and the secretions of interferon-γ （IFN-γ）, interleukin-17a （IL-17a）, and interleukin-22 （IL-22） were measured for all subsets. LX-2 cells were cultured in a serum-free medium and different concentrations of recombinant human IL-22 protein （25, 50, 100 ng/ml） were added; 24 hours later, the activation marker α-smooth muscle actin （α-SMA） was used to measure LX-2 activation. Fetal bovine serum with a volume fraction of 10% was used as a positive control. Enzyme-linked immunosorbent assay （chemiluminescence） was used to measure the concentrations of hyaluronic acid, type III precollagen, and type IV collagen in supernatant. A one-way analysis of variance, the non-parametric Mann-Whitney U test, and the non-parametric Kruskal-wallis H test were used for statistical analysis based on data type. Results Compared with the health control group, the liver cirrhosis groups with various causes had significant increases in peripheral Tcl, Th17, and Th22 cells. The percentage of Th17 ceils in the liver cirrhosis group was 1.64 times that in the control group （4.25%±2.45% vs 2.59%±1.36%, P 〈 0.05）, and the mean percentage of T