初发系统性红斑狼疮患者外周血CD4+调节性T细胞CD73表达

目的 研究CD73在初发活动性系统性红斑狼疮(SLE)患者外周血CD4+调节性T细胞的表达情况,探讨其在SLE发病中的作用.方法 采用流式细胞术检测29例初发未经治疗的活动期SLE患者(SLE组)和22例健康人(健康对照组)外周血CD4+CD25+CD73+T细胞百分率及CD4+CD73+、CD4+CDhi25、CD4+CD25+T细胞中叉头状转录因子3(FOXP3)蛋白表达,同时对CD73表达水平与SLE活动指标进行相关性分析.结果 SLE组患者外周血CD4+ CD25+ CD73+T细胞百分率低于健康对照组[(1.25±1. 32)%vs(2.35±1.09)%,P＜0.01].SLE组和健康正常对照组,CD73在C4+ CDhi25T细胞的表达水平[(29.05±12.53)%、(43.35±10.09)%]高于CD4+ CD25+T细胞[(17.48±6.92)%、(29.98±10.39)%,P＜0.001];FOXP3蛋白在CD4+ CD73+ T细胞[(65.36±14.40)%、(63.80±14.05)%]、CD4+ CDhi25 T细胞的表达水平[(67.30±13.04)%、(56.30±9.21)%]明显高于CD4+ CD25+ T细胞[(45.70±12.74)%、(43.98±5.17)%,P＜0.001],在CD4+ CD73-T细胞几乎不表达,而在CD4+ CD73+ T细胞、CD4+ CDhi25 T细胞中的表达差异无统计学意义(P值均大于0.05).CD73在CD4+ CD25+ T细胞的表达水平与SLE疾病活动指数、ESR、C反应蛋白、抗补体C1q、抗核小体抗体均无相关性(P＞0.05).结论 CD73可作为调节性T细胞新的表面标记,其在调节性T细胞中的异常表达可能参与SLE的发病机制.     

系统性红斑狼疮患者外周血CD4+CD25highFoxp3+调节性T细胞数量和Foxp3mRNA的表达水平与疾病活动性的相关性

目的 研究系统性红斑狼疮(SEE)患者CD4+CD25highFoxp3+调节性T细胞的数量及其功能基因Foxp3 mRNA的表达水平与SLE疾病活动性和肾脏损伤的相关性.方法 采用四色流式细胞术以Foxp3-异硫氰酸荧光素(FITC )/CD25-藻红蛋白/CD4-多甲藻叶绿素蛋白(PerCP)/CD3-藻蓝蛋白7抗体组合检测40名健康对照者及42例SLE患者外周血CD4+CD25highFoxp3+调节性T细胞的数量,实时荧光定量聚合酶链反应(PCR)检测特异性转录因子Foxp3 mRNA的表达水平,并分析其与SLE患者疾病活动指数(SLEDAI)、补体C3及血清抗双链DNA(dsDNA)抗体的关系.统计学方法采用t检验和Spearman相关分析.结果 活动期SLE患者外周血CD4+CD25highFoxp3+调节性T细胞数量显著低于健康对照组[(4±3)％与(7±4)％,P＜0.05],稳定期与健康对照组差异无统计学意义(P＞0.05);活动期SLE患者外周血CD4+CD25highFoxp3+调节性T细胞数量及CD4+CD25highFoxp3+调节性T细胞/CD4+比值显著低于稳定期患者[(4±3)％,(9±6)％与(5±4)％,(10±6)％,P均＜0.05];活动期SLE患者外周血Foxp3 mRNA的表达水平明显低于稳定期和对照组(P＜0.01,P＜0.05);SLE患者并发肾病组外周血CD4+CD25highFoxp3+调节性T细胞数量及CD4+CD25highFoxp3+调节性T细胞/CD4+比值显著低于SLE非肾病组(P＜0.05).相关分析显示,SLE患者外周血CD4+CD25highFoxp3+调节性T细胞数量与SLEDAI呈负相关(r=-0.5782,P＜0.05);CD4+CD25highFoxp3+调节性T细胞/CD4+比值与SLEDAI呈负相关(r=-0.4913,P＜0.05),与补体C3呈正相关(r=0.3687,P＜0.05);SLE患者外周血CD4+CD25highFoxp3+调节性T细胞数量与Foxp3 mRNA的表达水平呈正相关(r=0.6142,P＜0.0l).结论 SLE患者外周血CD4+CD25highFoxp3+调节性T细胞和Foxp3 mRNA的变化可能是导致SLE疾病发生和发展的关键因素之一,与疾病的活动性有密切关系.     

糖皮质激素对系统性红斑狼疮CD4+Foxp3+T细胞水平的影响

目的 探讨糖皮质激素治疗对系统性红斑狼疮(SLE)患者外周血CD4+ Foxp3+ T细胞水平的影响以及CD4+Foxp3+T细胞与SLE疾病活动的相关性.方法 采用流式细胞术检测26例SLE患者和5名正常人外周血CD4+Foxp3+T细胞百分率,Spearman相关分析法分析CD4+Foxp3+T细胞与SLE疾病活动指标及糖皮质激素的相关性.结果 活动期SLE患者外周血CD4+Foxp3+T细胞百分率(2.4±1.6)%低于正常人(3.3±0.8)%,但差异无统计学意义;非活动期SLE患者外周血CD4+Foxp3+T细胞百分率(3.3±0.7)%与正常人比较差异无统计学意义.活动期SLE患者经糖皮质激素治疗后外周血CD4+Foxp3+T细胞(6.1±3.5)%较治疗前(3.9±2.4)%升高(P＜0.05).SLE患者外周血CD4+Foxp3+T细胞水平与年龄、病程、红细胞沉降率、尿蛋白定量(24 h)、血清补体C3浓度、血清抗双链DNA(anti-dsDNA)抗体水平及SLE疾病活动指数(SLEDAI)无相关性,但与糖皮质激素每日用量呈显著正相关(r=0.51,P＜0.05).结论 SLE患者外周血CD4+Foxp3+T细胞水平不能作为狼疮活动指标.糖皮质激素治疗可上调SLE患者外周血CD4+Foxp3+T细胞水平,CD4+Foxp3+T细胞水平上调可能是糖皮质激素治疗SLE有效的机制之一.     

系统性红斑狼疮患者CD4~+CD25~+CD127~(low/-)调节性T细胞中Foxp3的表达及免疫抑制功能研究

目的 研究系统性红斑狼疮(SEE)外周血中调节性T细胞不同标志以及调节性T细胞在SLE发病中的作用;探讨CD127与Foxp3的相关性,明确CD127定义调节性T细胞的特异性;鉴定CD4~+CD25~+CD127~(low/-)T淋巴细胞免疫抑制功能.方法 ①采用四色直接荧光素标记法和多参数流式细胞术检测40例SLE患者(19例初发和21例缓解)及15名健康对照外周血CD4~+CD25~+T淋巴细胞、CD4~+CD25~+CD127~(low-)T淋巴细胞、CD4~+CD25~+Foxp3~+T淋巴细胞、CD4~+CD25~(high)T淋巴细胞、CD4~+CD25~(high)CD127~(low/-)T淋巴细胞、CD4~+CD25~(high)Foxp3~+T淋巴细胞以及CD4~+CD127~(low/-)Foxp3~+T淋巴细胞占CD4~+T淋巴细胞的比率,并且将7种调节性T细胞比率与外周血抗双链DNA(dsDNA)等抗体及SLE疾病活动指数(SLEDA1)评分等进行相关性分析.②以流式细胞分选术结合细胞培养技术,检测和分析3例SLE患者和4名健康人外周血中CD4~+CD25~+CD127~(low/-)调节性T细胞对CD4~+CD25~-效应性T细胞增殖的抑制作用.采用两样本均数的t检验,重复测量的方差分析,Pearson相关与Spearman相关分析进行统计学处理.结果 ①SLE患者组7种调节性T细胞比率分别为(6.1±1.7)%,(3.1±1.3)%,(2.1±1.0)%,(1.6±0.3)%,(0.97±0.28)%,(0.69±0.23)%和(0.71±0.35)%.与健康对照组比较:SLE患者组前6种调节性T细胞比率均低于健康对照组(P<0.05).②SLE患者组:CD4~+CD25~+Foxp3~+、CD4~+CD25~(high)Foxp3~+T淋巴细胞比率与IgA呈正相关;CD4~+CD25~(high)CD127~(low/-)T淋巴细胞比率与抗SSB抗体呈正相关.③SLE患者初发组和缓解组比较:SLE患者初发组7种调节性T细胞中除CD4~+CD127~(low/-)Foxp3~+T淋巴细胞比率外,其余均低于缓解组(P<0.05).④SLE患者初发组治疗前后比较:激素治疗前6种调节性T细胞比率均低于激素治疗后(P<0.05).⑤SLE患者初发组、缓解组和对照组中,CD4~+CD25~+T淋巴细胞及CD4~+CD25~(high)T淋巴细胞中Foxp3的表达与CD127低表达均呈正相关.⑥SLE患者、健康人CD4+CD25-效应性T细胞的体外增殖都可以被自身CD4~+CD25~+CD127~(low/-)调节性T细胞所抑制,但SLE患者的抑制率明显低于健康对照.结论 SLE的免疫异常可能与调节性T细胞的数量和功能缺陷有关;CD127可能代替Foxp3作为调节性T细胞特异性的表面标记物.     

L-选择素在系统性红斑狼疮患者外周血CD4~+CD25~+T淋巴细胞的表达及其与Foxp3的相关性

目的了解黏附分子L-选择素(L-selectin,CD62L)在系统性红斑狼疮(systemic lupus erythematosus,SLE)患者外周血CD4+CD25+T淋巴细胞的表达,及其与CD4+CD25+Treg细胞转录因子Foxp3的相关性。方法用流式细胞仪检测68例SLE患者外周血CD4+、CD4+CD25+T淋巴细胞CD62L和Foxp3的表达。根据SLEDAI评分,68例SLE患者中活动组36例,稳定组32例;健康对照组35人。结果活动组SLE患者外周血CD4+CD25+CD62L+T淋巴细胞为(1.71±1.60)%,低于对照组的(5.87±3.03)%(P0.01)和稳定组的(4.91±1.69)%(P0.01),差异具有显著性意义;且与SLEDAI呈负相关(r=-0.695,P=0.000),与补体C3水平呈正相关(r=0.522,P=0.000)。活动组SLE患者外周血CD4+CD25+CD62L+Foxp3+T淋巴细胞为(1.06±0.47)%,低于对照组的(3.17±0.87)%(P0.01)和稳定组的(3.46±1.15)%(P0.01),差异有显著性意义。CD4+T淋巴细胞CD62L的表达与CD4+CD25+T淋巴细胞CD62L和Foxp3的表达呈负相关(r=-0.689,P=0.000;r=-0.568,P=0.000);CD4+CD25+T淋巴细胞CD62L的表达与Foxp3的表达呈正相关(r=0.891,P=0.000)。结论CD62L在SLE患者外周血CD4+CD25+T的低表达及与Foxp3表达密切相关,可能在SLE发病中起着重要作用。     

HBV相关慢加急性肝衰竭患者外周血CD3+CD56+自然杀伤T淋巴细胞中TIM-3的表达及意义

目的探讨HBV相关慢加急性肝衰竭(HBV-ACLF)患者外周血自然杀伤T淋巴细胞(NKT细胞)数量及T淋巴细胞免疫球蛋白和黏蛋白结构域3(TIM-3)表达水平与患者肝损伤及预后的相关性。方法收集2016年9月-2018年6月在首都医科大学附属北京佑安医院接受治疗的43例HBV-ACLF患者和28例慢性乙型肝炎(CHB)患者的外周血单个核细胞,采用流式细胞技术检测患者外周血CD3+CD56+NKT细胞数量及其TIM-3表达水平。呈正态分布的计量资料2组间比较采用t检验;非正态分布的计量资料多组间比较采用Kruskal-Wallis H检验,2组间比较采用Mann-Whitney U检验;计数资料2组间比较采用χ2检验,相关性分析采用Pearson相关系数。结果 HBV-ACLF组ALT、AST、TBil、Cr、INR、HBV DNA、TBil/ALT比值及MELD评分等指标明显高于CHB组,Alb、PTA指标明显低于CHB组(P值均0. 05)。HBV-ACLF患者外周血CD3+CD56+NKT细胞数量为明显低于CHB组[(19. 13±13. 82)%vs (26. 75±11. 84)%,t=2. 401,P=0. 019],HBV-ACLF组CD3+CD56+NKT细胞TIM-3表达水平明显高于CHB组[5. 53%(2. 95%～10. 20%) vs 1. 59%(0. 91%～2. 70%),Z=-5. 260,P 0. 001]。CD3+CD56+NKT细胞TIM-3表达水平与ALT、AST、MELD评分、INR呈正相关(r值分别为0. 637、0. 414、0. 355、0. 335,P值分别为0. 000 1、0. 006、0. 021、0. 031),与PTA%呈负相关(r=-0. 313,P=0. 043)。43例HBV-ACLF患者中,早期患者为12例、中期患者为21例、晚期患者为10例,3组间CD3+CD56+NKT细胞数量比较差异均无统计学意义(P值均 0. 05),但呈增高趋势。中、晚期HBV-ACLF患者CD3+CD56+NKT细胞TIM-3表达水平分别为6. 50%(3. 16%～11. 45%)、8. 56%(4. 00%～10. 93%),均明显高于早期2. 58%(1. 92%～6. 02%)(Z值分别为-2. 284、-2. 641,P值均0. 05)。43例患者28 d存活组CD3+CD56+NKT细胞TIM-3表达水平明显低于28 d肝移植/死亡组[2. 98%(1. 94%～6. 88%) vs 8. 56%(4. 27%～11. 43%),Z=-2. 831,P=0. 005]。结论 HBV-ACLF患者外周血CD3+CD56+NKT细胞TIM-3表达水平增高,与患者肝损伤程度及预后相关。     

系统性红斑狼疮患者外周血淋巴细胞穿孔素表达及糖皮质激素对穿孔素的影响

目的 研究系统性红斑狼疮(SLE)患者外周血淋巴细胞穿孔素(PF)表达与分布,PF水平与疾病活动的相关性,探讨PF在SLE发病机制中的作用,观察糖皮质激素治疗对PF表达的影响.方法 SLE活动期患者16例,健康对照者12名.用流式细胞仪检测外周血淋巴细胞PF、CD3、CD4或CD8表达,用直线相关分析PF水平与SLE疾病活动性评分(SLEDAI)的相关性;比较糖皮质激素治疗前后PF水平变化.结果 与健康对照组比较,SLE活动期患者组外周血PF+淋巴细胞的百分率显著增高(P<0.01),PF的平均荧光强度(PFMFI)增高(P<0.01),PF+CD8+CD3+淋巴细胞的百分率明显增加(P<0.01),差异均有统计学意义.SLE活动期患者外周血PP淋巴细胞百分率与SLEDAI呈正相关(r=0.673).经过短期大剂量糖皮质激素治疗,SLE活动期患者外周血淋巴细胞PF+细胞百分率和PFMFI都显著下降(P<0.05),差异有统计学意义.结论 SLE活动期患者PF高表达,主要分布于CD8+CD3+细胞;PF水平与SLE活动程度呈正相关;短期糖皮质激素治疗能够减少患者体内PF表达.     

CD4+CD25-Foxp3+T淋巴细胞的表型鉴定及其在初发系统性红斑狼疮中的意义

目的 对初发系统性红斑狼疮(SLE)患者外周血异常表达CD4+CD25-Foxp3+T淋巴细胞进行表型鉴定,并探讨其临床意义.方法 对初发SLE患者外周血CD4+T淋巴细胞进行细胞表面分子[CD25、CD127、CCR4、糖皮质激素诱导的肿瘤坏死因子受体(GITR)、细胞毒T淋巴细胞相关抗原4(CT-LA-4)]和胞内分子(Foxp3)标染,流式细胞仪检测,并研究CD4+各细胞亚群与狼疮肾炎和疾病活动度(SLEDAI)相关性.结果 SLE患者外周血CD4+CD25-Foxp3+T淋巴细胞表面 GITR、CTLA-4和CCR4表达率与活化T淋巴细胞(CD4+CD25+Foxp3-)相比差异无统计学意义(P均>0.05),而显著低于调节性T淋巴细胞(CD4+CD25+Foxp3+)(P均<0.01);CD4+Foxp3+CD25high,CD4+Foxp3+CD25low和CD4+Foxp3+CD25-细胞中CD127low-百分率分别为(93.8±,3.5)%,(93.7±2.3)%,(92.0±2.1)%,三者之间差异无统计学意义(P>0.05);在CD4+细胞亚群中,当CD127low-时,Foxp3+在CD25high,CD25low和CD25-中表达率分别为 (91.4±2.6)%,(71.9±3.3)%,(9.0±2.2)%,三者之间差异均有统计学意义(P<0.01);SLE患者外周血CD4+CCR4+CD25highT淋巴细胞百分率与SLEDAI呈显著负相关(r=-0.695,P<0.001),狼疮肾炎患者(1.10±0.17)%显著低于SLE无肾炎组[(1.61±0.23)%,P<0.01]和健康对照组[(1.75±0.10)%,P<0.01];狼疮肾炎患者外周血CD4+ CCR4+CD25low-T淋巴细胞百分率显著高于健康对照组[(11.5 ±2.3)%与(8.0±1.0)%,P<0.01)].结论 初发SLE中异常升高的CD4+CD25-Foxp3+T淋巴细胞的表型类似早期活化效应T淋巴细胞.可以用CD4+CD25highCD127low-T淋巴细胞替选CD4+CD25highFoxp3+调节性T淋巴细胞.CCR4+调节性T淋巴细胞可能参与狼疮肾炎发病.     

外周血CD+4CD+25T细胞表达在系统性红斑狼疮患者中的意义

目的了解系统性红斑狼疮(SLE)患者外周血CD4 CD2 5T细胞表达及其临床意义。方法用流式细胞仪检测53例SLE患者外周血CD4 CD2 5T细胞表达,根据CD25表达荧光强度>100者为CD4 CD25brightT细胞,并与SLE活动指数评分(SLEDAI)、血清补体C3水平、抗双链DNA抗体、抗核抗体滴度进行相关分析。结果SLE患者外周血CD4 CD2 5T细胞表达率为(7·84±1·85)%,显著低于对照组(9·18±2·01)%(P<0·05),且活动期组[(6·72±1·16)%]较稳定期组更低[(8·57±1·91)%,P<0·01]。外周血CD4 CD25brightT细胞表达率在活动期组[(0·85±0·24)%]和稳定期组[(0·91±0·25)%]之间相比差异无统计学意义(P=0·686),但均低于对照组[(1·43±1·08)%,P<0·01]。随治疗后SLEDAI评分的下降,活动期组SLE患者外周血CD4 CD25brightT细胞表达率无明显变化。进一步分析外周血CD4 CD25brightT细胞表达率与SLEDAI评分、抗核抗体、抗双链DNA抗体滴度和血清C3水平的相关性,分别为ρ=-0·188,P=0·178;ρ=-0·216,P=0·121;ρ=0·082,P=0·560;ρ=0·010,P=0·944,差异均无统计学意义(P>0·05)。结论外周血CD4 CD2 5T细胞的减少可能与SLE的发病有关。     

系统性红斑狼疮患者外周血中γδT细胞的变化及临床意义

目的 探讨γδT细胞在系统性红斑狼疮(SLE)患者外周血中的变化及临床意义.方法 流式细胞术检测42例SLE患者、20名健康对照组外周血中CD3+γδ+T细胞百分率及绝对数;用膜联蛋白-Ⅴ/碘化丙啶法检测6例活动期SLE患者及6名正常对照组外周血中CD3+T细胞及γδ+T细胞的凋亡率.酶联免疫吸附试验( ELISA)法测定42例SLE患者血清抗核抗体和抗双链DNA(dsDNA)抗体水平.统计学方法采用t检验及Pearson相关分析.结果 活动期SLE患者外周血中T细胞百分率[(3.0±1.8)％]较非活动期[(5.3±3.0)％]及健康对照组[(6.8±2.8)％]均显著下降(t值分别为-3.071和-5.913,P均＜o.01);活动期SLE患者γδT细胞绝对数[(1.7±1.6)×107/L]明显低于非活动期[(5.3±3.6) ×107/L](t=-3.292,P＜0.01),且两者均低于健康对照组[(10.1 ±5.0) ×107/L](t值分别为-7.247和-2.905,P均＜0.01).活动期SLE患者γδT细胞绝对数与SLE疾病活动指数(SLEDAI)评分呈负相关(r=-0.365,P=0.047).SLE患者外周血中γδT细胞百分率(r=-0.336,P=0.030)及绝对数(r=0.410,P=0.007)均与红细胞沉降率( ESR)呈负相关,γδT细胞绝对数与血红蛋白呈正相关(r=0.409,P=0.007).此外,SLE患者外周血中γδT细胞凋亡率显著高于健康对照组及SLE患者CD3+T细胞凋亡率(t值分别为2.886和2.952,P均＜0.05).结论 SLE患者外周血中γδT细胞数量下降,与病情活动相关;γδT细胞数量下降的机制可能与凋亡增加有关.     

羟氯喹抑制紫外线诱导的系统性红斑狼疮CD4+T细胞白细胞介素-10和干扰素-γ的表达

目的 研究紫外线对系统性红斑狼疮(SLE)CD4+T细胞因子的影响和羟氯喹的抑制作用.方法 选择SLE 30例,健康对照10名.磁珠分选SLE患者和健康人的CD4+T细胞,紫外线311 nm窄谱中波紫外线暴露,加入羟氯喹共培养,酶联免疫吸附试验(ELISA)检测培养上清白细胞介素(IL)-10和干扰素-γ的表达水平.采用t检验进行统计学分析.结果 SLE患者CD4+T细胞IL-10表达高于健康对照[(27±4)和(18±3) pg/ml,P=0.011];经45、100 mJ/cm2紫外线暴露后,SLE活动患者CD4+T细胞IL-10表达升高[(27±4)和(77±42) pg/ml,(40±18)和(77±42) pg/ml,P=0.022,P=0.048],经100 mJ/cm2紫外线暴露后,活动患者CD4+T细胞IL-10表达高于稳定患者[(77±42)和(24±4)pg/ml,P=0.029];羟氯喹降低SLE活动患者CD4+T细胞IL-10和干扰素-γ表达[(2.6±4.0)和(17.9±2.3)pg/ml,P=0.018,P=-0.017)];羟氯喹降低经45,100 mJ/cm2紫外线暴露后SLE活动患者T细胞IL-10表达[(40±18)和(22±6)pg/ml,(77±42)和(21±5) pg/ml,P=0.037,P=0.04];羟氯喹降低经100 mJ/cm2紫外线暴露的SLE活动和稳定患者T细胞干扰素-γ表达[(18±3)和(13±14) pg/ml,(19±7)和(12±5) pg/ml,P=0.013,P=0.049].结论 紫外线加重SLE患者体内Th1/Th2细胞因子的比例失衡;羟氯喹抑制了紫外线诱发SLE患者干扰素-γ和IL-10的表达.

Abstract：

Objective To explore the role of hydroxychloroquine (HCQ) in ultraviolet B (UVB)- induced expression of interleukin (IL)-10 and interferon (IFN)-γ from CD4+T cells in patients with systemic lupus erythematosus (SLE). Methods Thirty patients with SLE and 10 healthy controls were enrolled in the study. CD4+ T cells were isolated using magnetic beads from SLE patients and healthy controls. HCQ was added in culture media before and after irradiation with UVB 311 nm narrow band ultraviolet B (NB-UVB). The levels of IL-10 and IFN-γ in the supernatant were detected with enzyme-linked immunosorbent (ELISA). Comparisons between groups were performed by t-test. Results The level of IL-10 was higher in SLE patients [(27±4) pg/ml] than that in healthy controls [(18±3) pg/ml, P=0.011]. After exposure of CD4+T cells to UVB in 45 or 100 mJ/cm2 dosages, the level of IL-10 was increased significantly in patients with active disease (P=0.022, P=0.048). After exposure of CD4+T cells to UVB in 100 mJ/cm2 dosages, the levels of IL-10 was higher in patients with active disease [(77±42) pg/ml] than patients with stable disease [(24± 4) pg/ml, P=0.029]. When CD4+ T cell were cultured with HCQ, IL-10 and IFN-γ levels in patients with active disease [(2.6±4.0), (17.5±2.3) pg/ml] were decreased significantly (P=0.018, P=0.017). HCQ reversed UVB-induced IL-10 expression in active SLE patients after exposure of CD4+T cells to UVB in 45 or 100 mJ/cm2 dosages (P=0.037, P=0.04). HCQ also reversed UVB-induced IFN-7 expression in active SLE patients and stable SLE patients after exposure to CD4+T cells with UVB in 100 mJ/cm2 dosages (P=0.013, P= 0.049). Conclusion UVB can aggravate the imbalance of Th1 and Th2 cytokines. HCQ inhibits UVB-induced IL-10 and IFN-7 expression of CD4+T cells in patients with SLE, especially in patients with active disease.     

系统性红斑狼疮患者外周血CD4+CXCR5+滤泡辅助性T细胞样细胞的变化及糖皮质激素对其的影响

目的 研究系统性红斑狼疮(SLE)患者外周血CD4+CXCR5+T细胞占CD4+T细胞百分率以及糖皮质激素对其的影响,探讨其在SLE发病机制中的作用.方法 采用流式细胞术检测45例活动期、20例缓解期SLE患者及20名健康对照外周血中CD4+CXCR5+T细胞占CD4+T细胞的百分率,比较其在各组中的差异及糖皮质激素治疗对其的影响,同时检测各组中CD19+B细胞上CXCR5的表达.2组间比较用独立样本t检验,3组间比较采用多变量方差分析,与临床指标之间的相关性分析采用非参数的Spearman相关分析,治疗前后的差异用重复测量的方差分析.结果 ①SLE患者外周血CD4+CXCR5+T细胞占CD4+T细胞的比例高于健康对照组[(16±7)%与(12±3)%,P＜0.01],其中活动组[(18±7)%]高于健康对照组(P＜0.05),而缓解组[(11±4)%]和健康对照组之间差异无统计学意义(P＜0.05);狼疮肾炎组高于非狼疮肾炎组,但差异无统计学意义[(18±7)%与(14±7)%,P=0.05].②CD4+CXCR5+T细胞百分率与SLE疾病活动指数(SLEDAI)、抗核抗体滴度和红细胞沉降率(ESR)呈正相关,与补体C3呈负相关(P均＜0.05),与C反应蛋白、病程、免疫球蛋白无相关性(P＞0.05).抗双链DNA抗体升高组与正常组之间、抗sm抗体、抗SSMSSB抗体阴性组和阳性组之间差异无统计学意义(P均＞0.05).③活动期SLE患者CD19+B细胞上CXCR5的表达比例低于健康对照组[(85±11)%与(94±3)%,P＜0.05].④10例初发、未接受治疗的活动期患者在接受地塞米松(20 mg/d)治疗后第1、3、7天外周血中CD4+CXCR5+T细胞百分率均低于治疗前(P均＜0.05).治疗前后CD19+CXCR5+B细胞的百分率无变化(P均＞0.05).结论 外周血CD4+CXCR5+滤泡辅助性T细胞样细胞的异常町能参与SLE的发病.

Abstract：

Objective To investigate the frequencies of CD4+CXCR5+T cells in the CD4+T cells of peripheral blood of patients with systemic lupus erythematosus (SLE) and the effect of glucocorticoid on it.Methods Frequencies of CD4+CXCR5+T cell were analyzed by flow cytometry in 45 active,20 inactive SLE patients and 20 healthy controls.Differences between groups and the effect of glucocorticoid were analyzed.Meanwhile, the expression of CXCR5 on CDI9+B cells was analyzed. Independent sample t test was used for statistical analysis between twogroups, ANOVA was applied for data analysis between 3 groups,,nonparameterical Spearman's analysis was used for correlation analysis and repeated measurement ANOVA were used to compare the parameters before and after treatment. Results The percentage of CD4+CXCR5+ in CD4+T cells was increased in patients with SLE compared with healthy controls[(16±7)% vs (12±3)%, P＜0.01].It was increased in patients with active SLE [(18±7)%] compared with healthy controls (P＜0.05) but there was no significant difference between inactive SLE[(11±4)%] and healthy controls(P＞0.05). The percentage in patients with LN was higher than that in patients without LN, but without significant difference[(18±7)%vs (14±7)%, P=0.05 ]. The percentage of CD4+CXCR5+T cells was positively correlated with SLEDAI,the titer of ANA and level of ESR but negatively correlated with the level of C3 (P<0.05 for each).No correlation was found between duration and the levels of CRP and immunoglobulin.. The percentage in patients with high anti-dsDNA group was also higher than that of the low group, but no differences were found between anti-Sm antibody positive and negative groups neither between anti-SSA/SSB antibody positive and negative groups(P＞0.05 for each).The expression level of CXCR5 on CD19+B cells in active SLE patients was lower than that of healthy controls[(85±11)% vs (94±3)%, P＜0.05 ]. The percentages of CD4+CXCR5+T cells in 10 untreated active SLE patients were decreased at day 1,day 3 and day 7 after being treated with dexamethasone (20mg/d) when compared with those before the treatment (P＜0.05 for each), but the percentages of CD19+CXCR5+B cells had no significant change (P＞0.05 for each).Conclusion These results demonstrate that the abnormality of CD4+CXCR5+T cells may play an important role in the pathogenesis of SLE.     

干扰素诱导蛋白44基因与系统性红斑狼疮的相关性研究

目的 探讨系统性红斑狼疮(SLE)患者外周血白细胞干扰素诱导蛋白 44(IFI44)基因实时定量表达水平与SLE临床表现及病情活动的相关性.方法 收集100例SLE患者,40例非SLE其他自身免疫性疾病患者,40名健康对照人群的临床资料,抽提外周血总RNA并逆转录成eDNA,运用SYBRgreen dve Ⅰ实时定量聚合酶链反应(RT-PCR)检测患者和对照组的IFI44基因定量表达水平,并对其与各临床指标及病情活动度的相关性进行分析.数据分析采用方差分析,采用Pearson及Spearman检验进行相关性分析.结果 ①SLE患者组的IFI44拷贝数26.8±5.3明显高于非SLE组7.4±2.7(P=0.0012)和健康对照组5.2±2.0(P=0.005);而非SLE患者组与健康对照组差异无统计学意义.②SLE重度活动组患者IFI44表达量63.1±22.4明显高于非活动组9.2±1.8(P=0.000)和轻度活动组患者28.0±7.2(P=0.015).③SLE患者组的IFI44拷贝数与SLEDAI积分之间呈正相关(r=0.38,P=0.000),与尿蛋白定量(24 h)也呈正相关(r=0.42,P=0.000).结论 IFI44基因在SLE患者中有明显表达上调现象;检测IFI44的表达水平有助于判断SLE患者病情活动状况.

Abstract：

Objective To investigate the expression of interferon-induced protein 44 (IFI44) gene in the leukocytes of the peripheral blood samples from patients with systemic lupus erythematosus (SLE), and to evaluate the relationship between the expression level and disease activity. Methods Mononuclear cells in the peripheral blood samples from 100 SLE patients were compared with those of 40 disease controls and 40 healthy donors (HD) and the expression of the IFI44 was evaluated by quantitative real-time PCR.Comparisons between groups were performed with ANOVA, and the correlation analysis between the level of expression was higher in SLE patients than disease controls and healthy donors (26.8±5.3, 7.4±2.7, 5.2±2.0,respectively) (P=0.0012, P=0.005), but no difference was found between disease controls and healthy donors. Mild disease activity and the SLE patients with stable disease (63.1±22.4, 28.0±7.2, 9.2±1.8, respectively)and 24 hours urine protein level (r=0.42, P=0.000). Conclusion IFI44 is demonstrated to be highly expressed in SLE patients. The level of IFI44 may be a promising candidate biomarker for identifying SLE activity.     

脐带间充质干细胞对系统性红斑狼疮患者Th17细胞的调节

目的 探讨脐带间充质干细胞(UC-MSCs)体内外对系统性红斑狼疮(SLE)患者CD3+CD8-IL17A+T淋巴细胞及相关细胞因子的调节作用.方法 UC-MSCs移植治疗14例SLE患者,观察移植前后的临床表现及实验窜指标的变化.采用流式细胞术检测各时间点患者外周血CD3+D8-IL17A+T细胞百分率,酶联免疫吸附试验(ELISA)测定血浆白细胞介素(IL)-6、转化生长因子(TGF)-β、IL-17A、IL-22表达水平.10例SLE患者外周血单个核细胞(PBMC)分别与UC-MSCs按不同比例体外共培养72 h,流式细胞术检测PBMC中CD3+CD8-IL17A+T细胞百分率.采用配对t检验和独立样本t检验.结果 UC-MSCs移植后患者疾病活动指数评分(SLEDAI)在3个月(7.8±1.2,t=2.19)及6个月(6.9±0.9,t=4.2)均显著低于移植前(10.4±0.9,P<0.05);尿蛋白定量(24 h)移植后6个月显著下降[(1489±260)与(2454±322)mg,t=2.6,P<0.05];血清白蛋白水平在移植后1、3、6个月均显著高于移植前(P<0.05),补体C3移植后持续升高.UC-MSCs移植后1个月及6个月,患者外周血CD3+CD8-IL17A+T淋巴细胞百分率显著下降(P<0.05).UC-MSCs与SLE患者PBMC共培养可显著下调CD3+CD8-IL17A+T淋巴细胞百分率(P<0.05),但无剂量依赖性.UC-MSCs移植后患者血浆IL-6、TGF-β、IL-17A、IL-22水平与移植前比,差异无统计学意义(P>0.05).结论 UC-MSCs移植治疗SLE有效,移植后SLE患者PBMC中CD3+CD8-IL17A+T淋巴细胞水平显著下降.

Abstract：

Objective To investigate the regulatory effects of umbilical cord-derived mesenchymal stem cells (UC -MSCs) on Th17 cells and related cytokines in patients with systemic lupus erythematosus (SLE). Methods Human UC-MSCs were isolated and expanded and infused into fourteen SLE patients. Clinical changes were evaluated before and after transplantation by SLE disease activity index (SLEDAI), 24-hour urine protein, serum albumin and complement C3. The percentages of CD3 +CD8-IL17A + T cells in peripheral blood were detected by flow cytometry. Concentrations of plasma IL-6, TGF-β, IL-17A, IL-22were determined by enzyme-linked immunosorbent assay (ELISA). UC-MSCs and peripheral blood mononindependent samples t-test. Results SLEDAI scores decreased significantly at 3 month (7.8±1.2, t=2.19) and 6 month (6.9±0.9, t=4.2) after UC-MSCs transplantation than pre-transplantation level (10.4±0.9, P< 0.05). Twenty-four-hour proteinuria decreased significantly 6 months after MSCs infusion [(1489±260) mg vs (2454±322) mg, t=2.6, P<0.05]. Meanwhile, serum albumin and complement C3 levels had increased significantly since 1 month after transplantation (P<0.05). The percentages of peripheral blood CD3+CD8-IL17A+T cells decreased obviously in 1 week, 1 month and 6 months after UC-MSCs transplantation (all P<0.05). The coculture of UC-MSCs with PBMC from SLE patients resulted in a statistically significant reduction of CD3+CD8-IL17A+T cells percentage in PBMC (P<0.05), but was not in a dose dependent manner. No change of plasma IL+6, TGF-β, IL-17A and IL-22 levels was observed after UC-MSCs transplantation (P>0.05).Conclusion UC-MSCs transplantation down-regulates the percentages of CD3+CD8-IL17A+T cells in SLE patients, which may be one of the mechanisms for its therapeutic effect in refractory SLE.     

Diminished CD4+CD25+ T-lymphocytes in peripheral blood of patients with systemic lupus erythematosus

The exact aetio-pathogenesis of systemic lupus erythematosus (SLE) is still speculative, where dysregulation or depletion of CD4+CD25+ T lymphocytes is among the supposed mechanisms. In this study, we thought to investigate patients with SLE for percentages of CD4+CD25+ T cells in their peripheral blood and to correlate this with their disease activity scores. Twenty-five patients with SLE who fulfilled, at least, four of the revised Criteria of the American College of Rheumatology (ACR) and twenty healthy volunteers participated in this study. Activity of SLE was assessed by SLE disease activity index (SLEDAI) score. Percentages of CD4+CD25+ T-cells were determined by a flowcytometric technique, while recently activated T-cells were analysed by assaying the expression of the T-cell activation marker, CD69. A statistically significant (p = 0.003) reduction of percentage of CD4+CD25+ T cells was observed among patients (mean 7.16+/-4.53 %) when compared with control subjects (mean 11.36+/-4.50 %), while a non-significant (p = 0.475) low expression of CD69 on CD4+ T cells was observed between patients (mean 0.32+/-0.28 %) and control subjects (mean 0.32+/-0.38 %). In addition, no correlation could be detected between percentages of CD4+CD25+ T cells and SLEDAI scores among SLE patients (p=0.079). In conclusion, this study adds some evidence for the role of CD4+CD25+ T cells in the pathogenesis of SLE that may have some future therapeutic applications.