冠状动脉旁路移植术后Th17和Treg细胞及其相关因子在新发心房颤动患者外周血中的变化

目的探讨辅助性T细胞17(Th17),调节性T细胞(Treg)及相关因子对冠状动脉旁路移植术后患者新发心房颤动(POAF)的影响。方法选择2013年5月至2016年7月132例行冠状动脉旁路移植术的冠心病患者,其中男82例、女50例,年龄39~76岁。收集患者术后2 h内静脉血,检测外周血Th17细胞,Treg细胞及相关因子表达水平,并进行数据统计分析。结果 132例患者中术后新发心房颤动35例(POAF组),未发心房颤动97例(No POAF组)。相较于No POAF组,POAF组患者外周血中Th17细胞比例和Th17/Treg比值明显升高(P0.05),而Treg细胞比例差异无统计学意义(P0.05)。血清中Th17细胞相关细胞因子(IL-6,IL-8和IL-17)含量均较No POAF组显著升高(P0.05)。而Treg细胞相关细胞因子(IL-10和TGF-β)含量在两组患者血清中含量差异无统计学意义(P0.05)。结论 POAF患者体内存在Th17/Treg平衡紊乱,调节Th17/Treg细胞平衡紊乱或可减少POAF的发生。     

强直性脊柱炎患者骨髓间充质干细胞的生物学及免疫学特性

目的:探讨强直性脊柱炎(AS)患者骨髓间充质干细胞(BMSCs)的生物学及免疫学特性,为进一步阐明AS的发病机制和寻找新的治疗靶点提供理论依据。方法 :选取37例活动期AS患者(AS组),男34例,女3例,平均年龄(24.3±5.4)岁,HLA-B27均为阳性;49例健康志愿者作为对照组(HD组),男43例,女6例,平均年龄(25.7±4.9)岁;其中HLA-B27阴性44例(HD1组),HLA-B27阳性5例(HD2组)。从每例受检者髂后上棘穿刺采集骨髓组织,分离BMSCs,培养扩增至第3代,以1×104/ml的浓度接种于96孔板中,100μl/孔,从第1天开始每日取3孔进行细胞计数,共计12d,绘制生长曲线;每日取3孔经MTT处理后测定吸光度值,绘制细胞活力曲线,观察BMSCs的生物学特性。使用流式细胞仪检测各受检者BMSCs的细胞表面表型。将第3代BMSCs细胞接种于U形底96孔板,培养4h后使用60Co照射30Gy;取健康志愿者外周血采用密度梯度法分离单个核细胞(PBMCs),加入细胞培养液,计数后按BMSCs∶PBMCs 1∶20、1∶10、1∶5、1∶2、1∶1的比例接种于已接种BMSCs的96孔板,共培养5d,观察双向混合淋巴细胞反应情况;同样获取PBMCs,计数后同样以5个比例接种于96孔板,加入植物血凝素(PHA)4μg/ml,充分接触后共培养5d,观察淋巴细胞增殖反应情况。对组间进行统计学比较。结果:AS组、HD1和HD2组第3代BMSCs体外培养1~12d时的增殖能力、细胞活力无显著性差异(P>0.05);三组细胞表面表型均为高水平表达CD105、CD73和CD90,不表达CD45、CD34、CD14和HLA-DR。HD1组和HD2组BMSCs与不同比例PBMCs共培养的双向混合淋巴细胞反应和PHA刺激的淋巴细胞增殖反应均无显著性差异(P>0.05);AS组与HD组比较有显著性差异(P<0.05)。结论:AS患者BMSCs的生物学特性无明显改变,但其免疫调节功能明显下降,其可能在AS的发病机制中扮演重要角色。     

特应性皮炎患者外周血Th17/Treg和IL-17、TGF-β的表达及与病情相关性研究

目的:探讨特应性皮炎(Atopic dermatitis,AD)患者外周血辅助性T细胞17(Th17)/调节性T细胞(Treg)平衡和白细胞介素-17(IL-17)、转化生长因子-β(TGF-β)的表达水平,并分析其与病情的相关性。方法:选取2015年3月-2019年6月在笔者医院确诊的AD患者126例作为观察组,再选取同期健康体检者60例作为对照组。分别检测两组静脉血中Th17、Treg细胞的百分比含量和IL-17、TGF-β水平。采用欧洲特应性皮炎评分标准(Scoring atopic dermatitis,SCORAD)对观察组病情进行评分,并分为以下三个亚组:轻度组(50例),中度组(42例),重度组(34例)。结果:观察组外周血中Th17、Th17/Treg和IL-17水平均高于对照组,Treg水平低于对照组,差异有统计学意义(P0.05)。与对照组比较,观察组TGF-β水平降低,但差异无统计学意义(P0.05)。重度组、中度组外周血中Th17、Th17/Treg和IL-17水平均高于轻度组,Treg、TGF-β水平均低于轻度组,差异有统计学意义(P0.05)。重度组外周血中Th17、Th17/Treg和IL-17水平均高于中度组,Treg、TGF-β水平低于中度组,差异有统计学意义(P0.05)。Pearson相关分析显示,患者外周血Th17/Treg、IL-17水平与其病情(SCORAD评分)之间呈正相关(r=0.930,0.893,P0.001)。患者TGF-β水平与其病情(SCORAD评分)之间呈负相关(r=-0.912,P0.001)。结论:AD患者外周血中Th17、Th17/Treg和IL-17水平均升高,Treg、TGF-β水平均降低,且患者Th17/Treg、IL-17、TGF-β水平均与其病情程度密切相关。因此,Th17/Treg细胞平衡在AD发病机制中发挥重要作用,在今后临床上可据此给予对应治疗。     

成人原发性肾病综合征中Th17/Treg免疫平衡及泼尼松对其影响的研究

目的研究成人原发性肾病综合征(primary nephrotic syndrome,PNS)患者外周血辅助性T细胞17(Th17)和调节性T细胞(Treg)的变化,探讨Th17/Treg免疫平衡在成人PNS发病中的作用以及泼尼松治疗成人PNS过程中对Th17/Treg免疫平衡的影响。方法选取初次就诊,经泼尼松治疗有效的患者65例,以年龄、性别匹配的25名健康体检者做为正常对照组,使用流式细胞术检测正常对照组和泼尼松治疗前、后患者外周血Th17细胞和Treg细胞比例,使用酶联免疫吸附试验(ELISA)法检测血清白细胞介素(interleukin,IL)-17和IL-10的水平,使用全自动生化分析仪检测24 h尿蛋白定量,分析Th17与其相关因子IL-17的相关性以及Treg与其相关因子IL-10的相关性,分析Th17/Treg比值与24 h尿蛋白定量的相关性。结果 PNS患者外周血Th17细胞比例为(2.44%±0.58%),较正常对照组的(0.93%±0.41%)明显升高,而PNS患者Treg细胞比例为(2.28%±0.63%),较正常对照组的(3.83%±1.06%)明显降低。经泼尼松治疗后,患者外周血Thl7细胞比例为(1.02%±0.51%),较治疗前明显降低,而Treg细胞比例为(3.69%±0.82%),较治疗前明显升高,差异均具有统计学意义(P0.05)。外周血Th17、Treg与相关细胞因子IL-17、IL-10的变化与两类细胞的变化一致。Th17/Treg细胞比值与24 h尿蛋白定量呈显著正相关(r=0.826,P0.01)。结论 Th17/Treg免疫失衡与成人PNS发生密切相关,泼尼松可能通过调节Th17/Treg免疫平衡影响成人PNS的发展,检测外周血Th17/Treg细胞比值的变化可做为成人PNS的辅助诊断指标以及泼尼松治疗成人PNS疗效评价指标。     

射频消融治疗对原发性肝癌患者T细胞免疫功能的影响

目的探讨射频消融(radio frequency ablation,RFA)前后原发性肝癌(primary hepatocellular carcinoma,PHC)患者T细胞亚群的变化。方法采用流式细胞术(flow cytometry,FCM)检测2011年1月~2013年6月40例PHC患者RFA术前24 h与术后1周、4周时外周血CD3~+、CD4~+、CD8~+、CD4~+/CD8~+、CD4~+CD25~+Treg细胞的变化;酶联免疫吸附试验(ELISA)检测血清Th1型细胞因子(IL-2、IFN-γ、TNF-α)含量。选择40例健康志愿者作为对照组。结果术前肝癌患者外周血T细胞亚群CD3~+、CD4~+、CD4~+/CD8~+明显低于健康对照组,CD8~+、Treg细胞明显高于健康对照组(P0.01);肝癌患者IL-2、IFN-γ、TNF-α明显低于健康对照组(P0.01)。术后1周,肝癌患者CD3~+、CD8~+、CD4~+/CD8~+与术前相比均无统计学意义(P0.05),术后4周CD3~+、CD4~+明显升高,CD8~+明显降低(P0.05);Treg细胞术后1周、4周与术前比较均明显降低(P0.01)。RFA治疗1周、4周IL-2、IFN-γ、TNF-α较治疗前均明显升高(P0.01)。结论射频消融可以改善肝癌患者短期内的T淋巴细胞功能,促进Th1型细胞因子分泌,从而对原发性肝癌患者的免疫状况产生有益的影响。     

Th17及Treg相关因子在反复种植失败患者种植窗口期子宫内膜中的表达

目的通过研究种植窗口期子宫内膜中Treg及Th17特异性转录因子及相关细胞因子表达情况,探讨反复种植失败患者子宫内膜中Treg/Th17平衡调节的可能机制。方法建立种植窗口期子宫内膜标本库。从标本库选择2013年10月至2014年12月收集的符合实验要求的反复种植失败患者(实验组)及首次移植妊娠患者(对照组)各20例,采用定量RT-PCR测量内膜标本中的Treg、Th17特异性表达因子Foxp3和ROR-γt及相关细胞因子IL-10、IL-17表达情况。结果在实验组和对照组子宫内膜上均可见Foxp3、ROR-γt及细胞因子IL-10、IL-17表达;与对照组比较,实验组Foxp3mRNA的相对表达量[(0.77±0.15)vs.(1.57±1.22)]及IL-10mRNA的相对表达量[(1.14±1.03)vs.(1.54±1.77)]均显著降低(P0.05),而ROR-γt mRNA的相对表达量[(0.92±0.39)vs.(0.55±0.15)]及IL-17mRNA的相对表达量[(0.95±0.31)vs.(0.49±0.21)]均显著升高(P0.05)。结论 IL-17、IL-10可能参与内膜中Th17/Treg平衡调节。     

Th17细胞和调节性T细胞在肝包虫病免疫逃避中的作用

目的 探讨辅助性T细胞17(Th17)和调节性T细胞(Treg细胞)在肝包虫病免疫逃避中的作用.方法 前瞻性分析2008年8月至2009年9月新疆医科大学第一附属医院74例受试者的临床资料,将74例受试者分为4组:健康对照组20例,肝囊型包虫病组21例,肝复发囊型包虫病组15例和肝泡型包虫病组18例.应用ELISA法检测各组受试者血清中Treg相关细胞因子转化生长因子(TGF-β1)和IL-10,Th17细胞相关细胞因子IL-17和IL-23的表达,并用单因素方差分析,两两比较采用LSD-t法和Pearson相关性检验分析其结果.结果 IL-17在健康对照组、肝囊型包虫病组、肝复发囊型包虫病组和肝泡型包虫病组的表达分别为(16±5)、(13±4)、(13±5)和(11±3)ng/L,4组比较,差异有统计学意义(F=6.35,P<0.05);而肝囊型包虫病组与肝复发囊型包虫病组比较,差异无统计学意义(t=0.22,P＞0.05).IL-23在健康对照组、肝囊型包虫病组、肝复发囊型包虫病组和肝泡型包虫病组的表达分别为(139±50)、(106±53)、(107±48)和(72±27)ng/L,4组比较,差异有统计学意义(F=6.74,P<0.05);而肝囊型包虫病组与肝复发囊型包虫病组比较,差异无统计学意义(t=0.02,P＞0.05).IL-10在健康对照组、肝囊型包虫病组、肝复发囊型包虫病组和肝泡型包虫病组的表达分别为(3.1±0.8)、(4.3±2.0)、(4.2±1.4)和(5.5±2.2)ng/L,4组比较,差异有统计学意义(F=9.78,P<0.05);而肝囊型包虫病组与肝复发囊型包虫病组比较,差异无统计学意义(t=0.14,P＞0.05);TGF-β1在健康对照组、肝囊型包虫病组、肝复发囊型包虫病组和肝泡型包虫病组的表达分别为(26±7)、(37±7)、(33±9)和(38±7)μg/L,4组比较,差异有统计学意义(F=6.73,P<0.05);而3种肝包虫病组比较,差异无统计学意义(t=0.56,1.81,P＞0.05).Th17/Treg(IL-17/IL-10)在健康对照组、肝囊型包虫病组、肝复发囊型包虫病组和肝泡型包虫病组的表达比例分别为5.7±2.6、3.6±1.5、3.4±1.9和2.1±0.7,4组比较,差异有统计学意义(F=13.76,P<0.05);而肝囊型包虫病组与肝复发囊型包虫病组比较,差异无统计学意义(t=0.23,P＞0.05).血清中,IL-17和TGF-β1呈负相关(r=-0.23,P<0.05);IL-17和IL-23呈正相关(r=0.70,P<0.05);IL-10和TGF-β1呈正相关(r=0.46,P<0.05).结论 Th17/Treg相关细胞因子平衡在肝泡型和肝囊型包虫病患者中明显向Treg应答偏倚,Th17/Treg相关细胞因子失衡可能参与肝包虫所致免疫逃避.     

肾移植受者外周血辅助性T淋巴细胞17、调节性T细胞表达及其平衡关系的研究

目的 探讨肾移植慢性排斥反应(CR)患者外周血辅助性T淋巴细胞(T helper,Th)17、调节性T细胞(regulatory T cells,Treg)、Th17/Treg变化及意义.方法 采用流式细胞术检测24例肾移植CR患者(CR组)和22名健康志愿者(正常对照组)外周血中的Th 17占CD4+T淋巴细胞的比例...     

mircoRNA-155表达量、CD4~+T细胞水平与幼儿湿疹的相关性分析

目的:探讨外周血中mircoRNA-155表达量、CD4~+T细胞水平与幼儿湿疹的关系。方法:选取2016年1月-2016年12月笔者科室诊治的120例湿疹患儿作为病例组,另选取同期60例健康幼儿作为对照组,对比两组外周血中mircoRNA-155表达量、CD4~+T细胞、嗜酸性粒细胞、血清免疫球蛋白E(IgE)水平,并探讨其关系。结果:病例组患儿的嗜酸性粒细胞、血清IgE、Treg、Th2及Th17水平均显著高于对照组,差异有统计学意义(P0.05);病例组患儿的Th1/Th2水平显著低于对照组,差异有统计学意义(P0.05)。慢性期、亚急性期、急性期患儿的嗜酸性粒细胞、血清IgE、Treg、Th2及Th17水平呈现逐渐升高的趋势,差异有统计学意义(P0.05)。慢性期、亚急性期、急性期患儿Th1/Th2呈现逐渐降低的趋势,差异有统计学意义(P0.05)。湿疹患儿Treg、Th2、Th17水平与嗜酸性粒细胞、IgE水平呈显著性正相关,差异有统计学意义(P0.05);湿疹患儿Th1/Th2水平与嗜酸性粒细胞、IgE水平呈显著性负相关,差异有统计学意义(P0.05)。结论:湿疹幼儿外周血中Th1/2细胞失衡与患儿病情变化具有相关性,mircoRNA-155表达变化不明显。     

大鼠肝移植免疫耐受过程Th细胞和Treg细胞因子及信号通路蛋白的变化研究

目的探讨大鼠肝移植免疫耐受过程中,辅助性T细胞(Th)和调节性T细胞(Treg)细胞因子及相关信号通路蛋白的变化与免疫耐受的关系。方法双袖套法建立原位肝移植模型,将大鼠分为3组:手术对照组(6只),假手术不进行肝移植;短期组(10只),术后存活10 d;耐受组(10只),术后存活100 d,移植肝功能恢复正常。检测各组大鼠肝功能指标如丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST),Th1细胞因子[干扰素(IFN)-γ、白细胞介素(IL)-2、肿瘤坏死因子(TNF)-α],Th2细胞因子(IL-4、IL-5、IL-6、IL-13),Th17细胞因子[粒细胞-巨噬细胞集落刺激因子(GM-CSF)、IL-17A],Treg细胞因子[IL-10、转化生长因子(TGF)-β、IL-12p]表达水平。对各组大鼠血清进行蛋白芯片检测。结果与手术对照组比较,短期组AST明显降低,ALT明显升高(P0.05)。而耐受组与手术对照组的AST、ALT水平比较,差异均无统计学意义(均为P0.05)。各组大鼠的肝组织中,与手术对照组比较,短期组Th1细胞因子IFN-γ、IL-2表达水平明显升高(均为P0.05);耐受组Th2细胞因子IL-4的表达水平明显低于手术对照组(P0.05);短期组Th2细胞因子IL-5、IL-6、IL-13的表达水平明显低于手术对照组(P0.05);耐受组IL-17A的表达水平明显高于手术对照组(P0.05)。耐受组IL-10、IL-12p的表达水平明显高于手术对照组(均为P0.05),短期组TGF-β的表达水平明显高于手术对照组(P0.05)。与手术对照组比较,短期组细胞间黏附分子(ICAM)-1、前血小板碱性蛋白(Ppbp)、神经纤毛蛋白(Neuropilin)-2、Notch-2蛋白的表达水平明显升高(均为P0.05);耐受组趋化因子配基17(CXCL17)、ICAM-1、Neuropilin-2蛋白的表达水平明显升高(均为P0.05),而B7-1蛋白的表达水平显著减低(P0.05)。结论在大鼠肝移植自然免疫耐受过程中,Treg类细胞因子(IL-10、TGF-β、IL-12p), IL-6, IL-17以及细胞表面的跨膜信号通路分子(ICAM-1、Neuropilin-2、B7-1蛋白)在其中起到了重要的作用。     

Th17/Treg cell balance in stable liver transplant recipients

BackgroundImmune monitoring of transplanted patients may provide a reliable basis for the individualization of immunosuppressive therapy. In addition, it might be applied for realizing the early and non-invasive diagnosis of acute allograft rejection.MethodsPercentages of TCD4 + IL-17+ (Th17) and TCD4 + CD25 + CD127^dim/− (Treg) cells, as well as serum levels of interleukin (IL)-17 and transforming growth factor (TGF)-β1, were evaluated in 30 stable patients using flow cytometry and ELISA techniques before and six months after liver transplantation. Besides, the same cells and cytokines were quantified in 10 recipients with acute allograft rejection.ResultsSix months post-transplant, the percentage of Th17 and Treg cells in the peripheral blood of stable liver transplant recipients reduced significantly, but the Th17/Treg ratios were comparable to the pre-transplant period (1.24 vs. 1.56); however, Th17/Treg ratios in the rejection group was significantly higher than in the stable recipients (4.06 vs. 1.56, P-value = 0.001). Stable patients showed decreased amounts of serum IL-17 which was remarkably lower than in the rejection group (P-value = 0.01). Moreover, there was a significant correlation between the serum level of IL-17 and the percentage of Th17 cells (P-value <0.001). Th17 frequency was negatively associated with the liver allograft function. Notably, TGF-β1 levels differed neither between pre-and post-transplant samplings nor between stable and rejection groups.ConclusionSix months after liver transplantation, the mean Th17/Treg ratio in stable recipients remained comparable to the pre-transplant values; however, it was significantly elevated in patients with acute allograft rejection, suggesting the Th17/Treg ratio as a probable predictor of acute rejection.     

Immunophenotyping of peripheral immunoregulatory as well as Th17A and Th22 cell subpopulations in kidney transplant recipients under belatacept or cyclosporine treatment

ObjectiveRegulatory Foxp3-expressing T cells (Tregs), IL-10-producing B cells (Bregs), and IDO-expressing dendritic cells (DCregs) downregulate inflammatory processes and induce peripheral tolerance, while Th17A and Th22 cell subpopulations are of proinflammatory nature. The aims of the study were to characterize and to enumerate peripheral Tregs, Bregs, and DCregs and Th17A and Th22 cell subpopulations in kidney transplant recipients (KTRs) under belatacept or cyclosporine treatment.MethodsForty-one KRT patients (30 under belatacept treatment and 11 under cyclosporine treatment) and 26 healthy donors (HDs) were included in the study. CD19⁺-expressing peripheral B lymphocytes were purified by positive selection. IL-10-producing B cells, CD4⁺/CD25^highFoxp3⁺, and CD8⁺/CD28^?Foxp3⁺ Tregs, CCR6⁺/CD123⁺/IDO⁺ DCs, as well as Th17A and Th22 cell subpopulations were quantitated by flow cytometry.ResultsOf the IL-10-producing Bregs, CD19⁺/CD24^high/CD38^high/CD5⁺, CD19⁺/CD24^high/CD38^high/CD10⁺, CD19⁺/CD24^high/CD38^high/CD20⁺, and CD19⁺/CD24^high/CD38^high/CD27^? had significant higher frequency in patients under belatacept treatment when compared with those under cyclosporine. Only CD19⁺/CD24^high/CD38^high/CD27⁺ and CD19⁺/CD24^high/CD38^high/CXCR7⁺ cells had significant higher frequency in patients under cycloporine treatment when compared to those under belatacept. The percentages of IDO-expressing pDC, CD4⁺/CD25^highFoxp3⁺, and CD8⁺/CD28^?Foxp3⁺ were significantly higher in the belatacept group when compared the cyclosporine one, while Th17A and Th22 cells had significant higher frequency in the latter group.ConclusionBelatacept seems to maintain and enhance, at least systemically, a tolerant profile to renal allograft in transplant recipients by means of higher circulatory frequencies of regulatory B, T and pDC subpopulations.     

Direct anti-proliferative effect of adipose-derived mesenchymal stem cells of ankylosing spondylitis patients on allogenic CD4+ cells

ObjectivesT-cell-mediated adaptive immunity contributes to the development and persistence of ankylosing spondylitis (AS). Mesenchymal stromal/stem cells (MSCs) have immunomodulatory potential and are able to inhibit T-cell proliferation, but their functionality in AS patients is relatively unknown. The aim of the study was to assess the direct anti-proliferative effects of MSCs isolated from subcutaneous abdominal adipose tissue of AS patients (AS/ASCs) on allogeneic T lymphocytes, using commercially available ASC lines from healthy donors (HD/ASCs) as a control.Material and methodsCD3+CD4+ T-cells were isolated from peripheral blood of healthy blood donors, activated with anti-CD3/CD28 beads, and co-cultured for 5 days with untreated and TNF+IFN-γ pre-stimulated HD/ASCs (5 cell lines) and AS/ASCs, obtained from 11 patients (6F/5M). The proliferative response of T-cells was analysed by flow cytometry, while the concentrations of kynurenines, prostaglandin E2 (PGE-2), interleukin 10 (IL-10), and interleukin 1 receptor antagonist (IL-1Ra) were measured spectrophotometrically or using a specific enzyme-linked immunosorbent assay (ELISA).ResultsHD/ASCs and AS/ASCs similarly reduced the T-cell proliferation response, i.e. the percentage of proliferating cells, the proliferation, and replication indices, and these effects were dependent mostly on soluble factors. In the co-cultures of activated CD4+ T-cells with HD/ASCs and AS/ASCs significant increases of kynurenines, PGE-2, and IL-1Ra, but not IL-10, production were observed. The release of these factors was dependent either on cell-to-cell contact (IL-10, IL-1Ra) or soluble factors (kynurenines, PGE-2). There was a moderate to strong negative correlation between T-cell proliferative response, and the concentrations of kynurenines, PGE-2, and IL-10, but not IL-1Ra. This association was more evident in the case of TI-treated AS/ASCs than HD/ASCs.ConclusionsAS/ASCs, similar to HD/ASCs, exert a direct effective anti-proliferative impact on CD4+ T cells, acting via soluble factors that are released in cell contact-dependent (IL-10) and independent (kynurenines, PGE-2) pathways. Thus, our results suggest that AS/ASCs are potentially useful for therapeutic application.     

Elevated RAP1A expression correlates with the severity of acute GVHD after umbilical cord blood transplantation

BackgroundAcute graft-versus-host disease (aGVHD) is a complication of allogeneic hematopoietic stem cell transplantation. Ras-related protein 1A (RAP1A) has been recently identified as a novel oncoprotein in several human malignancies. However, its specific role in aGVHD remains unclear.ObjectiveTo study the role of RAP1A in the pathogenesis of aGVHD.Material and methodsStudy participants included six patients with grade 2–4 aGVHD, 13 patients with grade 1 aGVHD, 11 patients without aGVHD, and 12 healthy people. The expression level of RAP1A in whole cells was detected by western blot and qRT-PCR. The proportions of CD4⁺CD25⁺FoxP3⁺ Treg cells (T regulatory cells) and the expression of RAP1A in Treg cells in peripheral blood mononuclear cells (PBMCs) were detected by flow cytometry and the levels of related cytokines in the serum was detected by cytometric bead array.ResultsWe found the level of RAP1A was higher in patients than in healthy individuals. A negative correlation was noted between RAP1A and the number of Treg cells. In addition, the level of IL-10 in patients with grade 2–4 aGVHD was significantly lower than that in healthy donors, however, the level of TNF-ɑ in patients with grade 2–4 aGVHD was higher. Furthermore, we found a negative correlation between levels of IL-10 and RAP1A, and a positive correlation between TNF-ɑ and RAP1A.ConclusionThe expression of RAP1A in patients with aGVHD was significantly increased, and shows potential as a target for the prevention and treatment of aGVHD.     

The imbalance between regulatory and IL-17-secreting CD4T cells in multiple-trauma rat

Background

It has been well recognised that a deficit of numbers and function of CD4⁺CD25⁺Foxp3⁺cells (Treg) is attributed to the development of auto-immune diseases, inflammatory diseases, tumour and rejection of transplanted tissue; however, there are controversial data regarding the suppressive effect of Treg cells on the T-cell response in auto-immune diseases. Additionally, interleukin-17 (IL-17)-producing cells (Th17) have a pro-inflammatory role. The balance between Th17 and Treg may be essential for maintaining immune homeostasis and has long been thought as one of the important factors in the development/prevention of auto-immune diseases, inflammatory diseases, tumour and rejection of transplanted tissue, but their role in multiple trauma remains unclear.

Objective

This study aims to investigate whether an imbalance of Treg and Th17 effector cells is characteristic of rats suffering from multiple trauma.

Methods and subjective

Sixty Sprague-Dawley (SD) rats were randomly divided into three groups. The control group (n = 20, group I) no received procedures (normal). The sham group (n = 20, group II) only received anaesthesia, cannulation and observation. The bilateral femoral shaft fractures with haemorrhagic shock groups (n = 20, group III). Rats in groups II and III were killed at the end of 4 h after models were established. Peripheral blood samples were collected for assessment of Treg cells, Th17 cells and cytokines (IL-17, IL-6, IL-2, transforming growth factor beta (TGF-β)) and intestine tissue was collected for intestine histological analysis.

Results

We observed decreased Treg/Th17 ratios in CD4⁺T cells in rats with multiple trauma and a strong inverse correlation with disease activity (intestinal histological scores).

Conclusion

We suggest a role for immune imbalance in the pathogenesis and development of multiple trauma. The alteration of the index of Treg/Th17 cells likely indicates the therapeutic response and progress in the clinic.     

Low-dose IL-2 expands CD4+ regulatory T cells with a suppressive function in vitro via the STAT5-dependent pathway in patients with chronic kidney diseases

Background: Patients with chronic kidney disease (CKD) often have CD4+ regulatory T cells (Tregs) dysfunction and chronic inflammation. We aim to investigate the effect, function, and related mechanism of low-dose IL-2 on CD4⁺ regulatory T cells expansion in vitro from patients with CKD.

Methods: A total of 148 newly diagnosed patients with CKD at Stage III and 35 healthy volunteer subjects were recruited into our studies. The number of peripheral Tregs in peripheral blood mononuclear cells isolated from CKD patients, which were characterized by FACS as CD4⁺CD25^hi and CD4⁺CD25⁺FoxP3⁺. The effect of low-dose IL-2 on expansion of Tregs, and the suppressive function of expanded Tregs were also analyzed by FACS. The levels of FoxP3 mRNA were detected by qRT-PCR. The activation of IL-2 induced Stat5 and blocking experiments were assessed by Western Blotting and FACS.

Results: We found that the frequency of peripheral Tregs from CKD patients was significantly lower than that in healthy volunteer subjects. We also showed that IL-2 selectively expanded CD4⁺CD25^hi and CD4⁺CD25⁺FoxP3⁺ regulatory T cells, and also upregulated the expression of FoxP3 mRNA. Our in vitro studies demonstrated that expanded CD4⁺ regulatory T cells from CKD patients suppressed proinflammatory Th1 and Th17 cell response. Furthermore, STAT5 activation is required for IL-2-induced expansion of regulatory T cells and expression of FoxP3 mRNA from CKD patients.

Conclusions: Our findings support the clinical Treg defects in CKD patients with glomerular diseases, and the rationale of evaluating low-dose IL-2 treatment for selectively modulating CD4⁺ Tregs.     

The prevalence of Th17 cells and FOXP3 regulate T cells (Treg) in children with primary nephrotic syndrome

The aim of this study was to investigate the prevalence of interleukin (IL)-17-producing CD4+ T cells (Th17) and regulatory T (Treg) cells in children with primary nephrotic syndrome. The study cohort consisted of 62 children who were randomly divided into control, primary nephrotic syndrome, and isolated hematuria groups. Flow cytometric analysis revealed the presence of Th17 cells in the peripheral blood mononuclear cells (PBMCs) of 35 children and Tregs in the PBMCs of all children. In addition, mRNA expression of Th17-related factors [IL-17, -23p19 and retinoid orphan nuclear receptor (RORc)] and the concentration of plasma inflammatory mediators such as IL-6 and IL-1β were consistently detected in all children. Protein expression of IL-17 and transforming growth factor-β1 were also detected in renal biopsy tissue and compared between different groups. Patients with PNS were found to have an increased number of Th17 cells and decreased numbers of Tregs in their PBMCs, and there was significant difference in the prevalence of Th17 and Tregs between the patients with PNS and those with isolated hematuria. Our data show that among our study cohort, there was a dynamic equilibrium between Th17 and Treg cells in children with PNS following the development of PNS with apparent renal tubular epithelial cell and interstitium lesions. The dynamic interaction between Th17 and Treg cells may be important in the development of PNS.