急性病毒性心肌炎患者CCL20对Th17细胞趋化性作用

目的:探讨CCL20与急性病毒性心肌炎(AVMC)患者Th17细胞趋化性的关系。方法:选择11例AVMC患者为研究对象,同时选取10例健康人作为对照。利用siRNA技术干预AVMC患者外周血巨噬细胞,将其分为A(不接受siRNA转染)、B(接受CCL20 siRNA转染)、C(阴性对照,接受scramble siRNA转染)亚组。应用ELISA法检测AVMC组患者血清CCL20及其巨噬细胞体外分泌CCL20的水平,趋化实验和流式细胞学结合观察体外不同水平CCL20对Th17细胞的趋化能力及表达CCL20受体CCR6的Th17细胞比例的影响,实时定量PCR法检测趋化板下室细胞表面CCR6mRNA的表达。结果:与健康对照组[(2.78±0.64)μg/L]相比,AVMC组患者体内血清CCL20水平[(11.36±2.45)μg/L]明显升高,P<0.05。体外实验中,B亚组巨噬细胞培养上清CCL20含量、趋化实验募集的Th17细胞比例、表达CCR6的Th17细胞比例和CCR6mRNA相对表达水平[(0.94±0.08)ng/L、(6.86±0.59)%、(2.56±0.78)%、(0.36±0.08)]均低于A亚...     

病毒性心肌炎小鼠辅助性T淋巴细胞17亚群的变化

目的 观察病毒性心肌炎小鼠不同时间点辅助性T淋巴细胞17(Th17)亚群的变化,探讨其在病毒性心肌炎发病中的作用.方法 用Balb/c小鼠建立柯萨奇病毒B3(CVB3)心肌炎小鼠模型(实验组),对照组注射等量磷酸盐缓冲液,在注射的第0、7、14、21、28和42天苏木精-伊红染色观察心肌病理改变并计算心肌病理积分,流式细胞仪检测小鼠脾脏Th17细胞亚群的比例(Th17/CD4+).结果 实验组7 d亚组小鼠心肌组织病理积分(1.8±0.5)高于0 d亚组,14 d亚组病理积分在实验组各亚组中最高(2.8±0.4),从21 d亚组起病理积分开始低于14 d亚组,实验组各亚组与对照组相应时点亚组比较差异均有统计学意义(P均<0.05).实验组7 d亚组小鼠脾Th17/CD4+[(2.23±0.89)%]高于实验组0 d亚组,28 d亚组脾Th17/CD4+最高[(5.00±0.81)%],42 d亚组脾Th17/CD4+[(2.35±0.35)%]低于实验组28 d亚组,与对照组相应时点亚组比较差异均有统计学意义(P均<0.05).结论 病毒性心肌炎小鼠中存在Th17细胞,其可能参与了病毒性心肌炎的炎性过程.     

CD5+B细胞与扩张型心肌病相关性研究

目的:通过对扩张型心肌病(DCM)患者体内CD5 B细胞及其功能的检测,探讨它在DCM发生发展中的作用。方法:选择DCM患者42例,20例健康者为对照。采用双色流式细胞术检测患者外周血淋巴细胞中CD5 B细胞(CD19 CD5 )数,ELISA法检测血清抗心肌自身抗体(AHA)以及体外CD5 B细胞分泌IL-10功能;根据彩色多普勒检测心脏射血分数(EF)。结果:与对照组[(5.39±1.46)%]相比,DCM患者CD5 B细胞均有明显增多,其中AHA阳性者为(13.49±3.15)%,AHA阴性者(14.06±5.23)%,AHA阳性和阴性者CD5 B细胞差异无统计学意义。对照组AHA检测均为阴性。DCM患者CD5 B细胞的增多与心脏EF值无相关性(r=-0.25,P>0.05),但其分泌IL-10功能与EF值负相关(r=-0.64,P<0.01)。结论:CD5 B细胞不仅参与AHA阳性也参与AHA阴性DCM发病过程;有活性的CD5 B细胞介导DCM心力衰竭的进程。     

辅助性T细胞17、辅助性T细胞17/辅助性T细胞1比例在急性冠状动脉综合征的变化及临床意义

目的探讨急性冠状动脉综合征(ACS)患者外周血辅助性T细胞(Th)17、Th17/Th1比例及血浆中白细胞介素17(IL-17)和IL-22的变化及意义。方法选择冠心病患者50例,其中急性心肌梗死(AMI)组20例,不稳定型心绞痛(UA)组15例,稳定型心绞痛(SA)组15例;健康查体者15例作为正常对照组。采用流式细胞分析法,检测各组外周血Th17、Th17/Th1细胞占CD4+T细胞比例。采用酶联免疫吸附法检测患者血浆IL-17和IL-22的浓度。结果AMI组和UA组外周血Th17细胞比例[(2.98%±1.01%)和(2.63%±0.61%)]、Th17/Th1细胞比例[(0.71%±0.35%)和(0.66%±0.31%)]均明显高于SA组及正常对照组(均P<0.05)。AMI组血浆IL-17(24.41±7.95 ng/L)和IL-22(34.18±7.04 ng/L)浓度均显著高于SA组和正常对照组(均P<0.05);UA组血浆IL-17浓度(22.86±8.62ng/L)显著高于正常对照组,血浆IL-22浓度(28.98±4.35 ng/L)显著低于AMI组而高于正常对照组。ACS组IL-22浓度与IL-17浓度呈显著正相关(r=0.422,P<0.01),且ACS组Th17细胞与IL-17(r=0.722,P<0.01)、IL-22(r=0.400,P<0.01)浓度均呈显著正相关。结论 Th17细胞可能参与动脉粥样斑块不稳定和ACS的发病。Th17细胞及其相关炎性因子IL-17、IL-22可作为诊断急性心血管事件发生的辅助检测指标。     

慢性特发性血小板减少性紫癜CD_4~+T细胞亚群变化的特点

目的 测定慢性特发性血小板减少性紫癜(ITP)患者CD_4~+T细胞亚群[Th1、Th2、Th17和调节性T细胞(Treg)]和相应血浆细胞因子水平,探索其在ITP发病机制中的作用.方法 将35例成人慢性ITP患者分为疾病活动组、未缓解组、缓解组,18例体检正常者设为对照组.应用流式细胞仪检测表达细胞内因子干扰素(IFN)γ~+、白细胞介素(IL)-4~+、IL-17~+细胞及表达CD_(25)~+Foxp3~+细胞分别占CD_4~+细胞的百分率.应用ELISA检测血浆中IFNγ、IL-4、IL-17和IL-10的水平,分析活动组患者血浆细胞因子水平与血小板、骨髓巨核细胞计数间的相关性.结果 Th1、Th17细胞百分率及Th1/Th17比例在各组间比较差异均无统计学意义;Th2细胞百分率:活动组[(1.01±0.88)%]、未缓解组[(1.22±1.04)%]与对照组[(1.86±0.59)%]比较明显下降(P<0.05);Th1/Th2比例:活动组(15.04±9.67)、未缓解组(11.65±9.32)与对照组(7.02±3.01)比较明显增高(P<0.05); Treg细胞百分率:活动组[(0.89±0.58)%]、未缓解组[(1.46±1.27)%]与对照组[(5.73±0.71)%]比较明显下降(P<0.01).IFNγ、IL-17水平在各组间比较差异无统计学意义;IL-4水平:活动组[(2.25±2.05)ng/L]、未缓解组[(2.33±2.14)ng/L]与对照组[(5.54±4.00)ng/L]比较明显下降(P<0.05);IL-10水平:活动组[(5.07±4.10)ng/L]、未缓解组[(5.66±4.35)ng/L]与对照组[(14.21 ±7.31)ng/L]比较明显下降(P<0.01).上述各参数(Th2、Treg细胞百分率、IL-4、IL-10水平)在缓解组与对照组间比较差异无统计学意义.活动组患者血浆IL-10水平与血小板计数呈正相关(r=0.16,P=0.03),与骨髓巨核细胞数间无相关性(r=0.41,P=0.06).结论 慢性ITP患者疾病活动组Th1/Th2比值升高,这是由Th2细胞的百分率下降所致.Treg细胞百分率下降可能与慢性ITP发病机制有关,但Th17细胞百分率可能与慢性ITP疾病状态无关.     

急性冠状动脉综合征患者外周血中辅助性T细胞亚群失衡促进血管内皮细胞的损伤作用

目的 分析急性冠状动脉综合征(ACS)和稳定性心绞痛(SAP)患者外周血中1型辅助性T细胞2型辅助性T细胞(Th1/Th2)的变化,探讨Th细胞在ACS中的变化及对血管内皮细胞损伤的作用.方法 收集40例ACS患者和18例SAP患者的外周血,检测外周血中Th1/Th2相关细胞因子的浓度,及不同细胞哑群对人脐静脉内皮细胞(HUVEC)的杀伤作用.结果 ACS组外周血中干扰素-γ(IFN-γ)和白细胞介素-2(IL-2)的浓度,均高于SAP组[IFN-γ:(131.2±42.2)ng/L比(47.6±20.2)ng/L,P<0.05;IL-2:(83.7±21.3)ns/L比(46.2 ±16.7)ng/L,P<0.05],ACS组外周血中IL-10浓度低于SAP组[(16.7±4.3)ng/L比(27.5±5.5)ns/L,P<0.05];ACS组患者的外周血单个核细胞(PBMC)对HUVEC的杀伤作用较SAP组明显增强(PBMC:28.84%±4.20%比20.28%±2.71%,P<0.05),而ACS组患者清除CD4+T细胞的PBMC[(CD4-)-PBMC]对HUVEC的杀伤作用与SAP组比较,差异无统计学意义[(CD4-)-PBMC:20.70%±3.26%比20.28%±2.71%,P>0.05].结论 ACS患者Th1亚群相关细胞凶子水平明显高于SAP患者,Th2亚群相父细胞因子水平显著低于SAP患者,Th1/Th2亚群失衡在PBMC对血管内皮的损伤中起促进作用,提示Th1/Th2亚群失衡可能参与了ACS的发生.     

Th17细胞和调节性T细胞在系统性红斑狼疮患者中的研究

目的 研究系统性红斑狼疮(SLE)患者外周血Th17细胞、调节性T细胞及联系两者的细胞因子白细胞介素(IL)-6的变化及其与病情活动的相关性,探讨Th17细胞、调节性T细胞在SLE发病机制中的作用.方法 收集103例SLE患者及28名健康者,SLE活动性的判断采用SLE疾病活动指数(SLEDAI)评分方法,其中非活动组(SLEDAI≤9分)患者37例,活动组(SLEDAI＞9分)患者66例.用四色分选流式细胞仪检测外周血Th17细胞、调节性T细胞百分数,用流式细胞仪微球捕获芯片技术(CBA)检测血清IL-6浓度,并分析其与病情活动的相关性.统计学处理采用t检验、秩和检验和Spearman相关分析.结果 ①SLE患者组Th17细胞[(1.2±1.1)%]、IL-6[(35±92)pg/ml]显著高于健康对照组[(0.6±0.4)%、(6±3)pg/ml],而调节性T细胞[(1.6±1.2)%]显著低于健康对照组[(2.6±1.8)%].②SLE活动组Th17细胞[(1.6±1.7)%]显著高于SLE非活动组[(1.0±0.7)%]与健康对照组(P均＜0.05),SLE活动组调节性T细胞[(1.5±1.3)%]显著低于SLE非活动组[(2.1±2.0)%]与健康对照组(P均＜0.05),SLE活动组Th17/调节性T细胞(0.68±0.34)显著高于SLE非活动组(0.24±0.20)与健康对照组(0.13±0.09,P均＜0.05),SLE活动组IL-6[(41±22) pg/ml]显著高于SLE非活动组[(32±28) pg/ml]与健康对照组(P均＜0.05);SLE非活动组与健康对照组之间Th17细胞、调节性T细胞、Th17/调节性T细胞、IL-6差异均无统计学意义(P＞0.05).③Th17细胞百分数与SLEDAI呈正相关,调节性T细胞百分数与SLEDAI呈负相关,Th17/调节性T细胞与SLEDAI呈正相关,IL-6浓度与SLEDAI评分之间呈正相关,Th17细胞与调节性T细胞呈负相关.结论 SLE患者外周血Th17细胞、调节性T细胞及IL-6浓度较健康对照组显著增高,并与疾病活动性密切相关,说明Th17细胞、调节性T细胞可能在SLE的发生、发展中起重要作用.

Abstract：

Objective To study the changes of Th17 cell, regulatory T cell (Treg) and interleukin (IL)-6 in the peripheral blood of patients with systemic lupus erythematosus (SLE) and their relationship with disease activity. Methods Percentage of Th17 and Treg in the peripheral blood of 103 patients with SLE and 28 healthy volunteers were detected by flow cytometry. The concentration of IL-6 in SLE patients and healthy volunteers was detected by cytometric bead array (CBA). The disease activity of SLE was measured by SLEDAI. SLE patients were divided into two groups: stable SLE (SLEDAI≤ 9, n=37) and active SLE (SLEDAI＞9, n= 66). The change of Th17, Treg, IL -6 and their relationship with disease activity were analyzed. Nonparamentric tests, t -test and spearman correlation were used for statistical analysis. Results The percentage of Th17 cells and the concentration of IL-6 in the peripheral blood in patients with SLE was higher than that in normal controls [respectively for (1.2±1.1)%, (35±92) pg/ml and (0.6±0.4)%, (6±3) pg/ml, P＜0.05]. However, the percentage of Treg in patients with SLE was lower than that in normal controls [respectively for (1.6±1.2)%,(2.6±1.8)%, P＜0.05]. The percentage of Th17, Th17/Treg IL-6 level in active SLE patients was higher than those in inactive SLE and those in normal controls (P＜0.05). However, the percentage of Treg in active SLE was lower than that in stable SLE patients and that in normal controls (P＜ 0.05). The percentage of Th17, Th17/Treg and concentration of IL-6 was positively correlated to disease activity(P＜0.05). But the percentage of Treg had negative correlation with the percentage of Th17 and disease activity (P＜0.05). Conclusion Th17, Treg and serum IL-6 in SLE patients are abnormal and they maybe contribute to the pathogenesis of SLE.     

原发性肝癌患者外周血CD4+T、CD8+T、Tc17、Th17和Treg淋巴细胞的变化及其意义*

目的 了解原发性肝癌(PLC)患者外周血CD4+T、CD8+T、Tc17、Th17和Treg淋巴细胞的变化。方法 2018年6月～2019年12月我院诊治的PLC患者83例(巴塞罗那临床肝癌分期A期25例,B期23例,C期18例,D期17例)和健康人35例,采用流式细胞技术检测外周血CD4+T、CD8+T及Tc17(CD8+ IL-17)、Th17(CD4+ IL-17)和CD4+CD25+CD45RA+ Treg淋巴细胞百分比。结果 PLC患者外周血CD4+T淋巴细胞百分比为(32.8±8.5)%,显著低于健康人[(43.3±7.4)%,P ＜0.05],CD4+/CD8 +细胞比值为(0.9±0.3),显著低于健康人[(1.2±0.1),P ＜0.05]; PLC患者外周血Treg细胞[(6.4±0.9)%对(3.0±0.1)%]、Th17细胞[(5.0±1.1)%对(3.1±1.5)%]及Tc17细胞[(2.3±0.4)%对(1.0±0.2)%],均较健康人显著升高(P＜0.05);BCLC C/D期患者外周血CD4+CD25+CD45RA+ Treg细胞百分比较A/B期患者显著升高[(7.6±0.4) %对(5.3±0.5)%,P＜0.001], Th17(CD4+ IL-17)和Tc17(CD8+ IL-17)细胞百分比较A/B期亦显著升高[分别为(6.9±1.1)%对(5.4±0.6)%,P ＜0.01和(2.8±0.6)%对(1.6±0.4)%,P＜0.01]。结论 PLC患者外周血淋巴细胞亚群出现异常改变,可能与肿瘤的分期密切相关,为从免疫学角度开展调节T淋巴细胞失衡的免疫治疗提供了理论依据。     

结核性胸腔积液中Th1/Th17细胞比例失衡及其机制

目的 探讨结核性胸腔积液患者外周血和胸腔积液中Th17细胞分布及其与Th1细胞的关系,以及结核性胸腔积液微环境中细胞因子对Th17细胞分化的影响.方法 2010年7月至2011年7月在武汉市结核病防治所住院的结核性胸腔积液患者30例,其中男12例,女18例;年龄16 ～63岁,平均41.2岁.采用流式细胞术检测患者胸腔积液和外周血Th1和Th17细胞亚群.分选初始T细胞诱导分化,以明确γ-干扰素、IL-11、IL-6和IL-12细胞因子对Th17细胞分化的影响.两组间比较采用t检验,多组间比较采用单因素方差分析,采用Pearson相关分析进行相关性检验.结果 结核性胸腔积液中Th1细胞占CD4+T细胞比例[(39±11)％]显著高于结核性胸膜炎[(8±3)％],差异有统计学意义(t=17.73,P＜0.05),Th17细胞占CD4+ T细胞比例[(2.8±0.9)％]显著高于结核性胸膜炎[(0.7±0.3)％],差异有统计学意义[t=14.78,P＜0.05].胸腔积液或外周血中Th1与Th17细胞亚群占CD4+T比例均呈正相关(r值分别为0.61和0.49,均P＜0.05).单用IL-1β、IL-6或二者联合应用可促进初始CD4+T细胞向Th17细胞方向分化,而γ-干扰素和IL-12能抑制Th17细胞分化,并可削弱IL-1β、IL-6对Th17细胞分化的促进作用.结论 结核性胸腔积液中存在Th1/Th17平衡,细胞因子IL-1β和IL-6可促进结核性胸腔积液中Th17细胞的分化.     

系统性红斑狼疮患者外周血白细胞介素-17与辅助T细胞17的检测及其意义

目的 研究系统性红斑狼疮(SLE)患者外周血白细胞介素(IL)-17蛋白和mRNA水平、辅助T细胞(Th17)细胞表达比例,探讨其临床意义.方法 用酶联免疫吸附试验检测SLE患者及对照者血浆中IL-17的蛋白水平;采用实时荧光定量反转录-聚合酶链式反应(RT-PCR)检测2组外周血中IL-17 mRNA表达水平;运用流式细胞术检测SLE患者外周血单个核细胞(PBMCs)中Th17细胞比例,进一步分析IL-17/Th17细胞与SLE临床实验室指标的相关性.计量资料组间比较采用t检验,相关性分析采用Spearman秩和相关分析.结果 SLE组患者血浆IL-17含量明显高于健康对照组,SLE组患者外周血IL-17 mRNA表达水平[(28.3±11.7)×10-5]高于对照组[(9.8±2.2)×10-5](P均＜0.01).SLE患者PBMCs中Th17细胞比例(2.5±1.5)%及IL-17荧光强度(1937±1022)显著高于对照组[(1.5±0.7)%,(1245±413)],且SLE活动期患者Th17细胞百分比高于非活动组,SLE肾病组Th17细胞比例较无肾病组明显升高(P均＜0.05).SLE患者血浆IL-17水平、Th17细胞数与SLE疾病活动性指数(SLEDAI)呈正相关(r=0.681,P＜0.01;r=0.426,P=0.034).结论 SLE患者体内IL-17蛋白分泌及基因表达水平存在明显异常,外周血Th17细胞比例亦显著升高,且与疾病活动性有明显相关,提示IL-17/Th17细胞可能在SLE发病中起着重要作用.     

Monoclonal anti-interleukin 23 reverses active colitis in a T cell-mediated model in mice

BACKGROUND & AIMS: Interleukin (IL)-23 supports a distinct lineage of T cells producing IL-17 (Th17) that can mediate chronic inflammation. This study was performed to define the role of IL-23 and Th17 cells in chronic colitis in mice. METHODS: Colitis was induced by transfer of a cecal bacterial antigen-specific C3H/HeJBir (C3Bir) CD4(+) T-cell line to C3H/HeSnJ SCID mice. Cytokines were measured by flow cytometry, enzyme-linked immunosorbent assay, and real-time polymerase chain reaction. Monoclonal anti-IL-23p19 was administered at the same time as or 4 weeks after pathogenic CD4 T-cell transfer. A histopathology colitis score was assessed in a blinded fashion. RESULTS: The pathogenic C3Bir CD4(+) T-cell line contained more cells producing IL-17 than those producing interferon-gamma and these were distinct subsets; after adoptive transfer to SCID recipients, Th17 cells were predominant in the lamina propria of mice with colitis. Bacteria-reactive CD4(+) Th1 and Th17 lines were generated. The Th17 cells induced marked inflammation in a dose-dependent manner. Even at a dose as low as 10(4) cells/mouse, Th17 cells induced more severe disease than Th1 cells did at 10(6) cells/mouse. Monoclonal anti-IL-23p19 prevented and treated active colitis, with down-regulation of a broad array of inflammatory cytokines and chemokines in the colon. Anti-IL-23p19 induced apoptosis in colitogenic Th17 cells in vitro and in vivo. CONCLUSIONS: Bacterial-reactive CD4(+) Th17 cells are potent effector cells in chronic colitis. Inhibition of IL-23p19 was effective in both prevention and treatment of active colitis. IL-23 is an attractive therapeutic target for inflammatory bowel disease.     

Salivary gland tissue expression of interleukin-23 and interleukin-17 in Sjögren's syndrome: findings in humans and mice

OBJECTIVE: Recently, the Th1/Th2 paradigm has been expanded by the discovery of Th17 cells, a subset of CD4+ memory T cells characterized by their unique ability to secrete interleukin-17 (IL-17) family cytokines. Importantly, Th17 cells appear to be intimately involved in autoimmunity. We undertook the present study to investigate whether the Th17/IL-23 system is up-regulated in Sj?gren's syndrome (SS). METHODS: Sera, saliva, and salivary glands from C57BL/6.NOD-Aec1Aec2 mice (a model for primary SS), as well as sera, saliva, and salivary gland biopsy specimens obtained from patients with primary SS, were evaluated for IL-17 and IL-23 expression by immunohistochemistry, real-time polymerase chain reaction, and the Luminex system. RESULTS: Immunohistochemical stainings of submandibular glands from C57BL/6.NOD-Aec1Aec2 mice and of salivary gland biopsy specimens from SS patients revealed strong positive staining for both IL-17 and IL-23 within lymphocytic foci and diffuse staining on epithelial tissues. Temporal expression of IL-17 and IL-23 in submandibular glands of C57BL/6.NOD-Aec1Aec2 mice correlated with expression of retinoic acid-related orphan receptor gammat, the Th17 cell master control gene. While IL-17 could not be detected in saliva from 4-20-week-old C57BL/6.NOD-Aec1Aec2 mice, this cytokine was present in the blood of mice up to age 16 weeks. This contrasted with sera and saliva from SS patients, in which IL-17 and IL-6 were present at varying levels. CONCLUSION: These results suggest that the Th17/IL-23 system is up-regulated in C57BL/6.NOD-Aec1Aec2 mice and SS patients at the time of disease. A correlation between up-regulated IL-17/IL-23 expression and specific clinical manifestations of SS has yet to be identified.     

Prostaglandin E2 exacerbates collagen-induced arthritis in mice through the inflammatory interleukin-23/interleukin-17 axis

OBJECTIVE: Recently, Th17 cells, a new subset of CD4+ T cells, emerged as major players in inflammation/autoimmunity. Maintenance of the Th17 phenotype requires interleukin-23 (IL-23), whereas the Th1-promoting cytokine IL-12p70 exerts a negative effect on Th17 cell differentiation. The lipid mediator prostaglandin E(2) (PGE(2)) acts primarily as a proinflammatory agent in autoimmune conditions, through mechanisms that remain to be elucidated. The aim of this study was to investigate whether PGE(2) released in inflammatory foci activates resident dendritic cells (DCs) to express IL-23 (at the expense of IL-12) and IL-6, resulting in a shift toward Th17 cell responses. METHODS: The effect of PGE(2) on IL-23 production by DCs and subsequent induction of T cell-derived IL-17 was assessed in vitro and in vivo. The effect of the stable PGE analog misoprostol was evaluated in a murine model of rheumatoid arthritis, in conjunction with IL-23 and IL-17 expression in affected joints and draining lymph nodes. RESULTS: In vivo administration of PGE(2) induced IL-23-dependent IL-17 production. Administration of misoprostol exacerbated collagen-induced arthritis (CIA). CIA exacerbation was associated with increased levels of IL-23p19/p40 messenger RNA and reduced expression of IL-12p35, and with increased levels of the proinflammatory cytokines IL-17, IL-1beta, IL-6, and tumor necrosis factor in the affected joint. Following ex vivo restimulation, draining lymph node cells from misoprostol-treated mice secreted higher levels of IL-17 and lower levels of interferon-gamma. CONCLUSION: Our results indicate that PGE(2) enhances DC-derived IL-6 production and induces a shift in the IL-23/IL-12 balance in favor of IL-23, resulting in increased IL-17 production, presumably through the amplification of self-reactive Th17 cells.     

Skewed distribution of Th17 lymphocytes in patients with Wegener's granulomatosis in remission

OBJECTIVE: The recently characterized interleukin-17 (IL-17)-producing T helper cell lineage (Th17), rather than the Th1 lineage, is involved in several autoimmune diseases. The possible role of Th17 cells in Wegener's granulomatosis (WG) has not yet been elucidated. We undertook this study to assess the distribution of Th1/Th2/Th17 cells and to investigate the presence of Th17 cells specific for the WG autoantigen proteinase 3 (PR3) in WG. METHODS: Peripheral blood from patients with WG in remission (n = 26) and healthy controls (n = 10) was stimulated in vitro with PR3 or with the control stimuli staphylococcal enterotoxin B (SEB), tetanus toxoid (TT), or phorbol myristate acetate/calcium ionophore, together with anti-CD28 and anti-CD49d. The frequencies of the various CD4+ T cell phenotypes responsive to stimuli were determined by 7-color flow cytometric detection of CD3, CD8, an early activation marker (CD69), and intracellular cytokines (IL-2, interferon-gamma [IFNgamma], IL-17, and IL-4). RESULTS: The percentage of CD69+,CD4+ T cells in patients with WG in remission was significantly decreased in response to PR3 and tended to be lower in response to other stimuli compared with the percentage in healthy controls. The percentages of Th17 cells (IL-4-,IL-17+,IFNgamma-) and Th2 cells (IL-4+,IL-17-,IFNgamma-) within the activated CD69+,CD4+ T cell population were significantly increased in patients with WG in remission, while no difference was found in Th1 cells (IL-4-,IL-17-,IFNgamma+) compared with the percentage in healthy controls. Increased percentages of Th17 cells in response to TT and SEB were found both in antineutrophil cytoplasmic antibody (ANCA)-positive and in ANCA-negative patients, while an increased frequency of PR3-specific Th17 cells was restricted to ANCA-positive patients. CONCLUSION: A skewed Th17 response found in ANCA-positive WG patients following stimulation with the autoantigen PR3 suggests that IL-17 is involved in disease pathogenesis and could constitute a new therapeutic target.     

Absence of donor Th17 leads to augmented Th1 differentiation and exacerbated acute graft-versus-host disease

Th17 is a newly identified T-cell lineage that secretes proinflammatory cytokine IL-17. Th17 cells have been shown to play a critical role in mediating autoimmune diseases such as EAE, colitis, and arthritis, but their role in the pathogenesis of graft-versus-host disease (GVHD) is still unknown. Here we showed that, in an acute GVHD model of C57BL/6 (H-2(b)) donor to BALB/c (H-2(d)) recipient, IL-17(-/-) donor T cells manifested an augmented Th1 differentiation and IFN-gamma production and induced exacerbated acute GVHD. Severe tissue damage mediated by IL-17(-/-) donor T cells was associated with increased Th1 infiltration, up-regulation of chemokine receptors by donor T cells, and enhanced tissue expression of inflammatory chemokines. Administration of recombinant IL-17 and neutralizing IFN-gamma in the recipients given IL-17(-/-) donor cells ameliorated the acute GVHD. Furthermore, the regulation of Th1 differentiation by IL-17 or Th17 may be through its influence on host DCs. Our results indicate that donor Th17 cells can down-regulate Th1 differentiation and ameliorate acute GVHD in allogeneic recipients, and that treatments neutralizing proinflammatory cytokine IL-17 may augment acute GVHD as well as other inflammatory autoimmune diseases.     

外源性细胞因子对柯萨奇病毒性心肌炎小鼠脾Th17细胞的影响

目的:观察转化生长因子(TGF)-β、白细胞介素(IL)-6和IL-23对柯萨奇病毒性心肌炎(VMC)小鼠脾中Th17细胞分化增殖的影响。方法:以6周龄雄性Balb/c小鼠腹腔注射柯萨奇病毒液建立VMC模型,1周后磁珠分离脾脏CD4+T淋巴细胞,分成对照组、TGF-β+IL-6组和TGF-β+IL-6+IL-23组进行体外培养。5d后应用流式细胞术测定刺激后Th17细胞的百分比;逆转录-聚合酶链式反应测定细胞中IL-17mRNA的表达;ELISA测定细胞培养上清液IL-17的水平。结果:与对照组比较,TGF-β+IL-6组Th17细胞数稍增加[(1.64±0.43)%∶(1.96±0.35)%],IL-17mRNA表达水平及细胞培养上清液IL-17浓度稍增加,但差异无统计学意义(P>0.05)。而TGF-β+IL-6+IL-23组Th17细胞数显著增加(4.08±0.77)%、IL-17mRNA表达水平及细胞培养上清液IL-17浓度均明显高于前2组,差异有统计学意义(P<0.05)。结论:外源性TGF-β联合IL-6对VMC小鼠脾中已分化的Th17细胞无增殖作用,而添加外源性IL-23则能显著增加Th17细胞数及其效应分子IL-17的表达。     

Mesenchymal stem cells improve the outcomes of liver recipients via regulating CD4+ T helper cytokines in rats

BACKGROUND:Bone marrow mesenchymal stem cells(BMMSCs) exert immunosuppressive activities in transplantation.This study aimed to determine whether BMMSCs reduce acute rejection and improve outcomes of liver transplantation in rats.METHODS:Orthotopic liver transplantation from Lewis to Brown Norway rats was performed,which was followed by the infusion of BMMSCs through the penile superficial dorsal vein.Normal saline infusion was used as a control.Animals were sacrificed at 0,24,72,or 168 hours after BMMSCs infusion.Liver grafts,and recipient serum and spleen tissues were obtained.Histopathology,apoptosis,serum liver enzymes,serum cytokines,and circulating regulatory T(Treg),Th1,Th2 and Th17 cells were assessed at each time point.RESULTS:BMMSCs significantly attenuated acute rejection and improved the survival rate of allogeneic liver transplantation recipients.Liver enzymes and liver apoptosis were significantly alleviated.The levels of the Th1/Th2 ratio-associated cytokines such as IL-2 and IFN-γ were significantly reduced and IL-10 was significantly increased.The levels of the Th17/Tregs axis-associated cytokines such as IL-6,IL-17,IL-23,and TNF-α were significantly reduced,whereas TGF-β concentration was significantly increased.Moreover,flow cytometry analysis showed that the infusion of BMMSCs significantly increased Th2 and Treg cells and decreased Th1 and Th17 cells.CONCLUSION:BMMSCs had immunomodulatory effects,attenuated acute rejection and improved outcomes of allogeneic liver transplantation in rats by regulating the levels of cytokines associated with Th1/Th2 and Th17/Treg ratios.