门静脉高压症脾亢患者外周血CD4+CD25+CD127low/-调节性T细胞含量与免疫功能研究

目的 研究门静脉高压症(PHT)脾亢患者外周血CD4+CD25+CD127low/-调节性T细胞(Treg)与CD3+、CD4+、CD8+T细胞的表达变化.探讨脾脏免疫功能改变及其可能的调节机制.方法 PHT脾亢患者20例(脾亢组),健康成人18例(对照组),采用流式细胞术检测外周血中CD4+CD25+CD127low/-Treg及CD3+、CD4+、CD8+T细胞的表达变化.结果 脾亢组外周血中CD4+CD25+CD127low/-Treg占CD4+T细胞的比例为(5.3±3.0)%,明显高于对照组的(2.5±0.9)%(P<0.01).脾亢组CD3+、CD4+、CD8+T细胞含量均低于对照组(P<0.05).CD4+CD25+CD127low/-Treg的含量与CD3+T细胞比例呈负相关(r=-0.630,P<0.01),与CD4+T细胞比例呈负相关(r=-0.561,P<0.05).结论 PHT脾功能亢进患者CD4+CD25+CD127low/-调节性T细胞表达量增多,可能对脾脏免疫功能的调节有重要作用.     

门静脉高压症脾功能亢进患者脾脏和外周血中CD4+CD25+CD1271ow/-调节性T细胞表达的研究

本研究通过流式细胞术检测门静脉高压症( portal hypertansion,PHT)脾亢患者外周血及脾脏组织中CD4+CD25+ CD127low/-调节性T细胞(CD4+ CD25+ CD127low/-Treg)的表达情况,探讨PHT脾亢患者脾脏免疫功能变化的可能机制,从而为PHT脾亢患者脾脏免疫功能的评估、具体手术方式的选择提供理论参考.     

肝癌病人外周血中CD4+CD25+FOXP3+调节性T淋巴细胞表达水平的临床研究

目的 了解CD4+CD25+FOXP3+T调节性T细胞在肝癌病人外周血中的表达水平并探讨其临床意义.方法 应用流式细胞术测定18例肝癌病人外周血CD4+CD25+FOXP3+调节性T细胞占CD4+T淋巴细胞百分比,并与26例l临床对照者和24例健康对照者进行比较.结果 肝癌病人外周血中CD4+CD25+T细胞占CD4+T细胞百分比(4.25%±3.98%)明显高于临床对照组(1.34%±1.14%)及健康对照组(1.29%±0.95%)(P＜0.01),而两个对照组之间并无显著性差异(P＞0.05).CD4+CD25+FOXP3+T细胞在肝癌病人外周血所占CD4+T细胞比率(2.94%±0.91%)也较临床对照组(0.76%±0.34%)及健康对照组(0.81%±0.29%)显著升高(P＜0.001),且升高幅度强于CD4+CD25+T细胞水平,两个对照组之间并无显著性差异(P＞0.05).结论 CD4+CD25+FOXP3+T细胞是更为准确的调节性T细胞,其在肝癌病人外周血中表达水平明显升高,检测CD4+CD25+FOXP3+T水平对肝癌的预防治疗具有重要临床意义.     

乙型病毒性肝炎肝硬化门静脉高压症患者脾切除后CD4＋CD25＋CD127low/-调节性T细胞的变化及意义

目的 观察乙型病毒性肝炎肝硬化门静脉高压症患者行脾切除术前后外周血中CD4＋CD25＋ CD127low/-调节性T细胞（Treg）的变化,探讨门静脉高压症患者行脾切除术对机体免疫功能的影响.方法 回顾性分析2012年5月至2013年5月河北医科大学第三医院收治的20例乙型病毒性肝炎肝硬化门静脉高压症合并脾功能亢进患者的临床资料,通过流式细胞仪分析乙型病毒性肝炎肝硬化门静脉高压症合并脾功能亢进患者（门静脉高压症组）脾切除术前1d、术后1周、1个月、3个月和来自河北医科大学第三医院的10例健康体检者（对照组）的外周血中CD4＋ CD25＋ CD127low/-Treg含量,分析Treg的变化对全身免疫系统造成的影响.组问比较采用t检验,手术前后数据比较采用重复测量方差分析.结果 门静脉高压症组患者术前和对照组受试者CD4＋ CD25＋ CD127low/-Treg比例分别为5.1％±3.5％和1.4％±0.2％,两组比较,差异有统计学意义（t=2.573,P＜0.05）.门静脉高压症组患者术后1周、1个月、3个月CD4＋CD25＋ CD127low/-Treg比例分别为9.2％±2.7％、5.6％±1.7％、2.5％±2.1％,其中术后1周与术前比较,差异有统计学意义（F=9.814,P＜0.05）;而术后3个月与术前比较,差异无统计学意义（F=2.364,P＞0.05）.结论 脾切除术后短期Treg水平明显升高,随时间延长逐渐下降.从Treg方面表明脾切除对机体免疫系统影响较小.     

Ⅲ型前列腺炎患者外周血中CD4+CD25+Treg及前列腺液中MCP-1的检测及意义

目的 探讨Ⅲ型前列腺炎/慢性骨盆底疼痛综合征(CAP/CPPS)患者外周血中CD4+CD25+调节性T细胞占CD4+T细胞的比率以及检测其前列腺液(EPS)中单核细胞趋化蛋白-1(MCP-1)的水平,分析各检测指标与临床症状的相关性.方法 采用流式细胞仪检测48例CAP/CPPS患者和10例正常对照者外周血CD4+CD25+Treg占CD4+T淋巴细胞的百分比;ELISA法检测两组受试者EPS中MOP-1水平.结果 CAP/CPPS组外周血中CD4+T细胞及CD4+CD25highTreg/CD4+T细胞(28.12±4.32)%,(3.99±0.61)%与对照组(28.29±4.30)%(3.96±0.66)%相比;差异无统计学意义,P>0.05.Ⅲ a组外周血中CD4+T细胞及CD4+CD25highTreg/CD4+T细胞(28.33±4.35)%,(3.98±0.60)%与Ⅲb组(27.91±4.26)%(4.01±0.62)%相比;P>0.05.Ⅲ型组CD4+CD25+Treg/CD4+T细胞(6.48±1.34)%,低于对照组(14.66±2.16)%;P<0.01.CAP/CPPS组外周血中CD4+T细胞以及CD4+CD25hignTreg/CD4+T细胞与患者慢性前列腺炎症状指数评分(CPSI)均无相关性(P>0.05):CD4+CD25+Treg/CD4+T细胞与患者疼痛评分呈负相关(r=-0.702,P<0.05).CAP/CPPS组外周血中CD4+CD25+Treg/CD4+T细胞与EPS中MCP-1水平呈负相关(r=-0.682,p>0.05).CAP/CPPS患者前列腺液中MCP-1(0.45±0.09)ng/ml较对照组(0.18±0.02)ng/ml显著升高;P<0.01.Ⅲ a组EPS中MCP-1水平(0.54±0.02)ng/ml较Ⅲb组(0.35±0.02)ng/ml显著升高;P<0.01.CAP/CPPS组EPS中MCP-1水平与NIH-CPSI呈正相关(r=0.716,P<0.01),且与患者疼痛评分明显相关(r=0.875,P<0.01),CAP/CPPS患者前列腺液中MCP-1水平与EPS中白细胞数呈正相关(r=0.898,P<0.01).结论 Ⅲ型前列腺炎患者外周血中CD4+CD25+Treg数量表达下调,导致患者自身免疫反应增强,可能是CAP/CPPS的发病机制之一;MCP-1在CAP/CPPS的发病过程中起重要作用,并且与临床症状密切相关,MCP-1可能成为CAP/CPPS临床诊断的一个指标.     

OX40信号调节抑制小鼠CD4+CD25+调节性T淋巴细胞Foxp3的表达

目的 探讨共刺激信号OX40对体外诱导的小鼠CD4+ CD25+适应性调节性T淋巴细胞(iTreg)的Foxp3表达的影响.方法 制备C57BL/6小鼠淋巴细胞悬液,经免疫磁珠法分选,获得CD4+ CD25-静息T淋巴细胞,与抗CD3单克隆抗体、抗CD28单克隆抗体、转化生长因子β1、白细胞介素2共孵育,诱导产生Foxp3+ iTreg.在此基础上,于培养体系中加入OX40激动型抗体及其对照抗体,利用流式细胞仪分析研究OX40信号刺激对iTreg Foxp3表达的影响.结果 C57BL/6小鼠淋巴结中CD4+ CD25+天然调节性T淋巴细胞(Treg)比例为(5.0±0.4)％,体外诱导培养的CD4+CD25+ Treg比例为(71.8±13.4)％,其中Foxp3阳性表达占(74.9±1.9)％.OX40激动型抗体组CD4+ CD25+ Treg细胞比例为(80.0±1.6)％,其中Foxp3表达水平为(59.2±0.7)％;OX40激动型抗体对照抗体组CD4+ CD25+ Treg细胞比例为(86.0±1.4)％,其中Foxp3表达水平为(70.0±0.8)％,两组间差异有统计学意义(P＜0.05).结论 静息T淋巴细胞可以在体外诱导培养获得高纯度iTreg;OX40信号刺激可以显著抑制CD25+ iTreg细胞Foxp3的表达.     

类风湿性关节炎患者外周血环瓜氨酸肽抗体与CD4+CD25+调节性T细胞检测的意义

目的 探讨类风湿性关节炎(RA)患者外周血环瓜氨酸肽(CCP)抗体、CD4+ CD25+调节性T细胞检测的意义.方法 54名符合1987年美国风湿病协会(ACR)修订的RA诊断标准的患者为实验组,20名正常体检健康人员做为对照组.用酶联免疫吸附试验(EUSA)检测血清CCP抗体,用流式细胞仪测定外周血CD4+ CD25+调节性T细胞水平,RA组患者均行双手和双足X线检查评价骨关节侵蚀情况.结果 RA组CCP抗体的阳性率为66.7％,较对照组明显增高(P＜0.01);RA组及对照组CD4+ CD25+调节性T细胞表达水平分别为(3.41±1.83)％、(4.23±2.16)％,两组差异有统计学意义(P＜0.05);RA患者CCP抗体阳性组与阴性组CD4+ CD25+调节性T细胞表达水平分别为(3.17±1.57)％、(3.59±1.84)％,差异无统计学意义(P＞0.05),两组骨关节侵蚀发生率分别为72.2％、16.7％,差异有统计学意义(P＜0.05),CCP抗体阳性组骨关节侵蚀率与CD4+ CD25+调节性细胞水平无明显相关(r=-0.126,P＞0.05).结论 CCP抗体可以作为RA的重要实验室检测指标;CCP抗体阳性RA患者较阴性患者更易发生骨关节侵蚀,但骨关节侵蚀的发生可能与CD4+ CD25+调节性T细胞的表达水平降低无关.     

乙型肝炎患者外周血中CD4+CD25+调节性T细胞的分子表达研究

目的:分析乙型肝炎患者外周血Tregs水平并探讨其在HBV感染者中发病机制的作用.方法:采用流式细胞术检测不同类型乙型肝炎患者及健康对照组外周血Tregs的水平及应用荧光定量PCR技术测定其血清中HBvDNA载量.结果:①慢性乙型肝炎组(CHB)Tregs表达率较急性乙型肝炎组(AHB)有明显差异(P＜0.05),与无症状携带者组及健康对照组则有显著差异(P均＜0.01);②AHB组较健康对照组的稍低,但无统计学意义(P>0.05),且明显低于CHB组(P＜0.01)；③无症状携带者组与健康对照组相比无显著差异(P＞0.05).④CHB组外周血HBVDNA载量较其他三组明显偏低(P＜0.01)；AHB组与无症状携带者组、健康对照组相比,差异无统计学意义(P＞0.05).⑤不同类型HBV感染患者外周血Tregs表达率与HBVDNA载量呈正相关.结论:①外周血Tregs表达的水平与乙肝患者病情轻重有一定关系,它在乙型肝炎持续感染中可能发挥了重要作用.②不同类型HBV感染患者外周血Tregs表达率与HBVDNA载量呈正相关.     

肾病综合征患者外周血中CD4+CD25+调节性T细胞的表达和意义

CD4+CD25+调节性T细胞(CD4+CD25+Treg)数量或功能的降低可导致多种自身免疫性疾病发生.肾病综合征(NS)与T细胞免疫功能紊乱密切相关.本研究采用CD4、CD25和CD127三标法检测初诊及不同治疗阶段NS患者外周血中该群细胞,观察活化效应细胞/调节性细胞(CD127hi/CD127low)的改变,从而探讨CD4+CD25+Treg与NS发生、发展之间的关系.     

胃癌患者CD4+CD25+Foxp3+调节性T细胞与TGF-β1检测的意义

目的:探讨胃癌患者外周血CD4+CD25+Foxp3+调节性T细胞(CD4+CD25+Foxp3+Tergs)以及血清TGF-β1水平的变化及意义。方法:检测并比较42例胃癌患者(胃癌组)与24例健康体检者(对照组)外周血CD4+CD25+Foxp3+Tergs与血清TGF-β1水平,分析两者水平与胃癌患者临床病理因素的关系。结果:胃癌组CD4+CD25+Foxp3+Tergs和TGF-β1水平均明显高于对照组(均P<0.05)。胃癌患者外周血CD4+CD25+Foxp3+Tergs水平与TNM分期、淋巴结转移有关(均P<0.05),而血清TGF-β1的表达水平与TNM分期、分化程度、淋巴结转移有关(均P<0.05);胃癌组患者外周血内CD4+CD25+Foxp3+Tergs的表达水平与血清内TGF-β1的表达水平呈明显正相关(r=0.801,P<0.05)。结论:胃癌患者外周血CD4+CD25+Foxp3+Tergs水平及血清TGF-β1水平升高,检测两者水平有助于患者病情与预后的判断。     

Acceleration of apoptosis in CD4+CD8+ thymocytes by rapamycin accompanied by increased CD4+CD25+ T cells in the periphery

BACKGROUND: Rapamycin (Rapa) is an immunosuppressant that is used in patients and animal models to control allograft rejection. Its mechanisms of action are not fully understood. In this article, the authors have investigated the effects of therapeutic doses of Rapa on both thymic and peripheral T-cell populations in the adult rat. METHODS: The therapeutic dosage of Rapa was optimized using cardiac transplantation between LEW and DA rats. Thymic morphology was assessed by hematoxylin-eosin staining. Flow cytometric analysis was performed to analyze T-cell phenotype and apoptosis. T-cell receptor (TCR)-mediated T-cell responsiveness was evaluated by 3[H]-thymidine deoxyribose incorporation. RESULTS: Rapa induced atrophy in the thymus but not in peripheral lymphoid organs. Moreover, fibrosis occurred in thymus that was long-lasting after Rapa withdrawal. In animals treated with Rapa, there was a significant reduction in CD4+CD8+ thymocytes caused by accelerated apoptosis, whereas CD4-CD8-, CD4+CD8-, and CD8+CD4- populations remained unaffected. In contrast, the cellularity of the periphery lymphoid organs was not altered. Within the CD4+ thymocyte population, CD4+CD25+ thymocytes were resistant to Rapa-accelerated apoptosis, and in the periphery, the ratio of CD4+CD25+ to CD4+CD25- T cells was increased. Notably, the peripheral CD4+CD25+ T cells were hyporesponsive to TCR-mediated activation. CONCLUSIONS: The resistance of the peripheral CD4+CD25+ T cells to Rapa treatment might contribute to its immunosuppressive action. The long-term effects of Rapa on thymus atrophy and thymocyte development requires consideration with respect to its clinical application.     

CD19+CD5+ B cells in primary IgA nephropathy

The source of IgA and the mechanism for deposition of IgA in the mesangium remain unknown for primary IgA nephropathy. Because CD19(+)CD5(+) B cells are important producers of IgA and contribute to several autoimmune diseases, they may play an important role in IgA nephropathy. In this study, flow cytometry, quantitative PCR, and confocal microscopy were used to assess the frequency, distribution, Ig production, CD phenotypes, cytokine production, and sensitivity to apoptosis of CD19(+)CD5(+) B cells in the peripheral blood, peritoneal fluid, and kidney biopsies of 36 patients with primary IgA nephropathy. All patients with IgA nephropathy were significantly more likely to have CD19(+)CD5(+) B cells in the peripheral blood, peritoneal fluid, and kidney biopsies than were five control subjects and 10 patients with active systemic lupus erythematosus. The 33 patients who had IgA nephropathy and responded to treatment demonstrated a significant decrease in CD19(+)CD5(+) B cells in the peripheral blood, peritoneal fluid, and kidney (all P < 0.01). In the three patients who had IgA nephropathy and did not respond to treatment, the frequency of CD19(+)CD5(+) B cells did not change. CD19(+)CD5(+) B cells isolated from patients with untreated IgA nephropathy expressed higher levels of IgA, produced more IFN-gamma, and were more resistant to CD95L-induced apoptosis than cells isolated from control subjects and patients with lupus; these properties reversed with effective treatment of IgA nephropathy. In conclusion, these results strongly suggest that CD19(+)CD5(+) B cells play a prominent role in the pathogenesis of primary IgA nephropathy.     

CD4+CD25+ cells regulate CD8 cell anergy in neonatal tolerant mice

BACKGROUND: Injection of neonatal BALB/c mice with semi-allogeneic splenocytes leads to antigen-specific tolerance lasting into adulthood. Tolerant mice accept A/J skin grafts and fail to generate CD8 cytotoxic T lymphocyte (CTL) activity against A/J targets. Anergic CD8 T cells are present in tolerant mice, and CD4 regulatory cells function to maintain CD8 cell anergy. METHODS: Neonatal BALB/c mice were injected with 108 live CAF, splenocytes, and mice were deemed tolerant by accepting A/J grafts over 40 days. CD8 cell proliferation was measured by in vitro incorporation of bromodeoxyuridine coupled with fluorescence-activated cell sorter analysis. Alloantigen-specific cytotoxicity was tested using 51Cr release assays of A/J or third-party targets. RESULTS: We demonstrate that A/J-specific anergic CD8 cells are present in neonatal primed mice that develop tolerance but not in neonatal primed mice that reject A/J skin grafts. Anergic CD8 cells show decreased proliferation and no CTL activity against A/J targets. Addition of interleukin-2 (IL-2) to unfractionated cultures fails to restore CTL activity against A/J targets. However, addition of IL-2 to CD4-depleted cultures restores A/J-specific CD8 CTL activity. Removal of CD4+/CD25+ cells, but not CD4+/CD25- cells, also restores CD8 CTL activity against A/J in the presence, but not the absence, of IL-2. Moreover, when added back into cultures, purified CD4+/CD25+ cells from tolerant mice inhibit the generation of CD8 CTL against A/J targets. CONCLUSION: These data indicate that CD8 anergy is associated with the state of tolerance, and that CD4+CD25+ cells from tolerant mice function to maintain A/J-specific CD8 cell anergy in vitro.     

肝癌、肝硬化组织中CD3、CD57、CD20细胞数量与临床病理表现及预后的关系

目的探讨CD3、CD57、CD20细胞在原发性肝细胞癌(HCC)、癌旁、肝硬化及正常肝组织中的数量及意义.方法HCC 60例,单纯性肝硬化62例,正常肝组织23例,以免疫组化SP法进行CD3、CD57、CD20染色,对阳性细胞数进行定量分析并与临床资料进行相关探讨.结果(1)各组CD3+细胞平均数从高到低为癌旁组织、癌组织、肝硬化组织、正常肝组织(P＜0.05);各组CD57+细胞平均数从高到低为癌组织、癌旁组织、正常肝组织、肝硬化组织(P ＜0.05);各组CD20+细胞平均数从高到低为癌组织、癌旁组织、肝硬化组织、正常肝组织(P ＜0.01).(2)HCC中CD3+细胞、CD57+细胞、CD20+细胞与组织学分级均无明显关系.(3)HCC中CD57+细胞和CD20+细胞随着临床分期的发展有下降的趋势(P ＜0.05);HCC中CD3+细胞平均数与临床TNM分期无关.(4)HCC中15月内有转移组的CD57+、CD3+细胞数均少于无转移组(P＜0.01).HCC患者15月内有无转移与HCC和癌旁组织中的B细胞分布均无关.结论临床上,随着HCC患者的病情恶化,CD3+、CD57+、CD20+细胞逐渐减少.CD3+、CD57+、CD20+细胞可成为反映机体抗肿瘤特异性细胞免疫状态和生物学行为及判断患者预后的重要指标.     

血清微环境对小鼠T细胞衰老的调节作用

目的：探讨血清微环境对小鼠T细胞衰老的调节作用。方法：分别取年老（12～14月龄）及年轻（1.5～2月龄）小鼠各10只,提取其脾脏淋巴细胞及血清,实验分4组。组I为年老鼠T淋巴细胞＋10%年轻鼠血清;组II为年老鼠T淋巴细胞＋10%年老鼠血清;组III为年轻鼠T淋巴细胞＋10%年轻鼠血清;组IV为年轻鼠T淋巴细胞＋10%年老鼠血清。培养48h后,经流式细胞术研究CD8＋CD28＋共表达率差异。结果：组I和组II T细胞表面的CD8＋CD28＋共表达率分别是（10.84±0.6841）%和（3.18±0.1789）%,组III和组IV T细胞表面的CD8＋CD28＋共表达分别是（12.5±0.9445）%和（8.36±0.2074）%。各组间对比有统计学差异（P〈0.05）结论：血清微环境具有调节小鼠T细胞衰老的作用,年轻鼠血清能使年老鼠的T细胞表面的CD8＋CD28＋共表达率提高,年老鼠的血清能使年轻鼠的T细胞表面的CD8＋CD28＋共表达率降低。     

CD4+CD25+ regulatory T cells mediate acquired transplant tolerance

The Holy Grail of clinical organ transplantation is the safe induction of allograft tolerance. Transplant tolerance has been successfully induced in animal models. Since T cells play a pivotal role in graft rejection, modulating T cell function has been the primary focus of studies aimed at inducing transplant tolerance. Rodent models of transplant tolerance induction include central deletion and peripheral mechanisms involving activation-induced cell death (AICD), anergy, immune deviation, and production of regulatory T cells. These mechanisms are not mutually exclusive. Although clonal deletion and anergy limit self-reactive T cells in the thymus, these mechanisms alone are not sufficient for controlling self-reactive T cells in the periphery. There is now evidence that the adult animal harbors two functionally distinct populations of CD4(+) T cells; one mediates autoimmune disease and the other dominantly inhibits it. The latter cells express CD4, CD25 and CTLA-4. These thymus-derived T cells have recently been shown to mediate the induction and maintenance of transplant tolerance. These CD4(+)CD25(+) T cells are similar in origin, phenotype, and function to those that maintain natural self-tolerance and T cell homeostasis in the periphery. Against this background, is it possible that alloantigen specific regulatory T cells might be generated and expanded ex vivo before organ transplantation and then infused to induce long-term tolerance, perhaps without the need for chronic immunosuppression?