系统性红斑狼疮患者外周血CD4+ CD8+和CD22+淋巴细胞活化分子CD69的表达

目的探讨系统性红斑狼疮(SLE)患者外周血淋巴细胞(PBL) T细胞(CD4+、CD8+)和B细胞(CD22+)活化分子CD69的表达.方法应用双染色流式细胞术检测CD4、CD8、和CD22细胞亚群CD69分子;在植物凝集素(PHA)刺激后20 h淋巴细胞亚群CD69分子的表达.结果①SLE患者PBMC的CD69分子活动期高于静止期(P＜0.001)和正常对照组(P＜0.01)的表达,SLE静止期患者与正常对照组CD69表达差异无显著性(P＞0.05).②进一步分析CD4+、CD8+和CD22+淋巴细胞亚群的CD69的表达,其中,SLE活动期患者CD4+细胞的CD69表达显著高于静止期(P＜0.001)和正常对照组(P＜0.01)的表达,SLE静止期患者与正常对照组CD69表达差异无显著性(P＞0.05);CD8+细胞活动期高于静止期患者(P＜0.05),其余组间差异无显著性(P＞0.05);CD22+B细胞各组间差异无显著性.③PHA刺激20 h后,CD4+、CD22+B细胞的CD69表达,活动期显著高于静止期患者和正常对照组(P＜0.01).结论 SLE患者外周血CD4+T细胞和CD22+B细胞存在着异常的活化,这种淋巴细胞的异常活化是SLE重要的发病机制之一.     

共刺激分子CD28 CD152在类风湿关节炎患者T细胞亚群上表达异常的研究

目的观察共刺激分子CD28,CD152在类风湿关节炎(RA)患者的T细胞亚群上的表达异常情况,探讨RA的发病机制及治疗手段.方法用流式细胞仪采用直接免疫荧光法测定39例RA患者和20名健康对照人外周血T细胞表面标志CD3,CD4,CD8的表达情况及CD28,CD152在CD4+T和CD8+T细胞上的表达.结果 RA患者CD3+CD4+细胞较正常对照组显著增高(P＜0.01),CD3+CD8+细胞较正常对照组显著降低(P＜0.05),CD4+T细胞上CD28的表达较对照组显著降低(P＜0.05),而CD8+T细胞上CD28的表达与对照组差异无显著性(P＞0.05);CD4+T和CD8+T细胞上CD152的表达都较对照组显著增高(P＜0.01).结论在RA患者的细胞免疫活化过程中首先表现为B7/CD28信号途径占优势,T细胞被激活,激活的T细胞大量分泌CD152,它与CD28竞争结合B7分子,CD152/B7途径转而占优势,下调或终止T细胞反应.同时CD28+细胞数目的减少或功能缺陷造成RA患者外周血单个核细胞凋亡加速,是诱发RA患者的局部病理损害的原因.阻断CD152和B7的相互作用可增强特异性T细胞应答,为RA的免疫学治疗提供理论依据.     

慢性乙型肝炎患者外周血T细胞亚群和共刺激分子的表达及意义

探讨慢性乙型肝炎患者外周血T细胞亚群及共刺激分子表达的临床价值。采用流式细胞术检测了 6 8例慢性乙型肝炎患者和 5 0例正常人外周血T细胞亚群及共刺激分子的表达。慢性乙型肝炎中、重型患者CD 4 细胞及CD4/CD8细胞比值明显低于正常组。三组患者CD2 8、CD 8CD 2 8表达水平均显著低于正常组 ,而CD80 、CD 8CD-2 8表达水平均显著高于正常组。CD2 8、CD80 、CD 8CD 2 8、CD 8CD-2 8的表达及CD4/CD8细胞比值在各型患者间均有明显差异 ,而CD3 、CD8表达水平无明显变化。T细胞亚群及共刺激分子的检测 ,对指导乙肝的临床治疗、判断预后有一定意义     

慢性乙型肝炎患者外周血CD8~+T细胞CD95表达的研究

目的检测慢性乙型肝炎患者外周血CD8+T细胞CD95分子的表达水平,了解CD95分子表达与炎症进展的关系。方法采集37例慢性乙型肝炎患者及健康对照者外周血,分离外周血单个核细胞(PBMCs),用流式细胞术检测外周血CD8+T细胞CD95分子的表达情况。结果高病毒载量组和低病毒载量组CD8+T细胞CD95的表达均显著高于健康对照组(P0.05);CD95的表达及CD8+T细胞频率之间存在负相关关系(r=-0.3486)。结论慢性乙型肝炎患者CD95的高表达可能促进了CD8+T细胞的凋亡,使CD8+T细胞的抗病毒作用减弱,可能与HBV持续感染、慢性乙型肝炎的形成机制有关。     

原发性胆汁性肝硬化T淋巴细胞亚群分析

目的 探讨原发性胆汁性肝硬化(PBC)患者的T淋巴细胞亚群及共刺激信号表达的特点及临床意义.方法 以未经治疗的98例PBC患者为研究组,性别、年龄匹配的健康人30名作为对照组.以流式细胞仪技术检测外周血淋巴细胞亚群以及T淋巴细胞表面的共刺激信号CD28.结果 PBC与对照组T细胞亚群差异有统计学意义:PBC组CD4+T淋巴细胞升高,CD8+T淋巴细胞下降,CD4+/CD8+比值上升(P<0.05);CD4+CD28-T细胞和CD8+CD28-T细胞明显增加(P<0.05).结论 PBC存在免疫调节功能的异常,其中CD28的表达明显减少;CD8+CD28-的细胞群可能在PBC中有一定的免疫调节作用.     

血液透析对尿毒症患者外周血T细胞亚群的影响

目的　探讨终末期尿毒症患者外周血T细胞亚群功能以及血液透析对其的影响。方法　应用流式细胞仪法检测 30例透析及未透析患者外周血T细胞表面的CD3、及其亚群CD4及CD8抗原表达。结果　尿毒症未透析组 (NHD组 )的外周血T细胞的CD3、CD4和CD8表达及CD4+ /CD8+ 比值均显著低于正常对照组 (P<0 0 5 ) ;透析组 (HD组 )与NHD组相比 ,其CD3、CD4和CD8值及CD4+ /CD8+ 比值均显著上升 (P <0 0 5 ) ,其中CD8值接近于正常对照组。结论　尿毒症患者的T细胞免疫功能明显失衡 ,血液透析疗法能够部分改善患者的T细胞亚群免疫功能。     

肺肿瘤患者围手术期CD28及T细胞亚群检测的临床意义

目的:检测肺肿瘤患者手术前后外周血CD28和T淋巴细胞亚群的表达变化并探讨其意义。方法:采用流式细胞术检测63例肺肿瘤患者手术前后外周血CD28及T淋巴细胞亚群的表达,并与健康对照组及肺良性肿瘤组作比较。结果:肺肿瘤组术前外周血中CD3+、CD4+、CD4+/CD8+、CD8+CD28+水平明显低于健康对照组和肺良性肿瘤组,而CD8+水平明显升高。肺肿瘤组CD8+表达水平随着临床分期的进展而有显著升高,CD3+、CD4+、CD4+/CD8+和CD8+CD28+水平随着临床分期的进展而有显著降低。不同病理分型的肺肿瘤患者之间外周血CD28及T细胞亚群水平无明显差异。肺肿瘤患者术后CD3+、CD4+、CD4+/CD8+和CD8+CD28+表达显著升高;CD8+的表达明显降低,但仍未恢复至健康水平。结论:肺肿瘤患者CD3+、CD4+、CD4+/CD8+和CD8+CD28+表达水平低下,从而影响B7-CD28共刺激通路,使T细胞不能有效的清除肿瘤,提示肺肿瘤患者机体免疫功能低下,施行手术切除肿瘤有助于改善患者的细胞免疫功能。     

多发性骨髓瘤患者T细胞共刺激分子表达和调节性T细胞检测及意义

目的通过检测多发性骨髓瘤患者外周血T细胞共刺激分子CD28表达及CD4^＋CD25^high调节性T细胞（Tregs）水平,探讨其免疫功能状态及临床意义。方法采用流式细胞术检测38例多发性骨髓瘤患者及20例健康志愿者外周血T细胞亚群cD。表达和CD4^＋CD25^high Tregs水平。分别采用透射免疫比浊法、溴甲酚绿法及速率法测定患者血清β2-微球蛋白（β2--MG）、白蛋白（Alb）及乳酸脱氢酶（LDH）水平。结果与正常对照组相比,多发性骨髓瘤患者外周血CD4＋T细胞百分率降低、CD8^＋T细胞百分率升高、CD4／CD8比值降低（P〈0．05或P〈0．01）,进一步研究发现CD4^＋和CD8^＋T细胞CD28表达均较对照组明显下降,而CD4^＋CD25^high Tregs百分率显著升高（P〈0．05,P〈0．01）。CD4^＋和CD8^＋T细胞CD28表达在进展期明显低于稳定期（P均〈0．01）,而CD4^＋CD25^high Tregs在稳定期和进展期间比较差异无统计学意义（P〉0．05）。多发性骨髓瘤患者CD4^＋T细胞CD28表达、CD8^＋T细胞C28表达均与其血清β2-MG水平呈负相关,而与Alb水平呈正相关（P〈0．05,P〈0．01）,CD4^＋CD25^high Tregs与Alb水平呈负相关（P〈0．05）。结论多发性骨髓瘤患者存在T细胞亚群共刺激分子CD28表达缺陷和CD4^＋CD25^high Tregs扩增,并与病情进展及预后存在一定相关性。     

CD178在肾综合征出血热患者外周血T细胞亚群表达的研究

为研究 CD1 78在肾综合征出血热 (HFRS)患者外周血 T细胞亚群的表达及意义 ,应用荧光抗体标记和流式细胞仪技术分析 CD1 78(Fas配体 )在 HFRS患者及健康人群 (对照组 )外周血 CD 4、CD 8T细胞表达的水平。结果 HFRS患者发热期组和多尿期组外周血 CD 4T淋巴细胞比例与正常对照组比较差异无显著性 ;而 CD 8T细胞明显增加 (P<0 .0 1) ;CD 4/CD 8比值明显下降 (P<0 .0 5 ) ;CD 4、CD1 78 T淋巴细胞和 CD 8、CD1 78 T淋巴细胞皆显著增高 (P<0 .0 5及 P<0 .0 0 1)。HFRS患者发热组和多尿组 CD 8T细胞的 CD1 78的表达率分别为 2 2 .18%和2 9.92 % ,而 CD 4T细胞的 CD1 78表达率分别为 4 .80 %和 5 .2 5 %。认为 HFRS的发病过程中 CD1 78主要表达于CD 8T淋巴细胞亚群 ,且发病早期和末期均有高表达     

EB病毒感染所致肝脏损伤患者病毒特异性CD4+T细胞和CD8+T细胞功能分析

目的分析EB病毒(EBV)感染者中发生肝脏损伤和未发生肝脏损伤患者病毒特异性CD4+T细胞和CD8+T细胞功能的差异。方法入组45例EBV感染者, 28例发生肝脏损伤,17例未发生肝脏损伤。分离外周血单个核细胞, 纯化CD4+T细胞和CD8+T细胞, 使用重组EBV核心抗原2(EBNA2)刺激培养96 h。细胞技术试剂盒法检测细胞增殖, 流式细胞术检测CD4+T细胞和CD8+T细胞比例。酶联免疫吸附试验检测CD4+T细胞分泌细胞因子和CD8+T细胞分泌毒性分子水平, 实时定量PCR法检测CD4+T细胞亚群转录因子mRNA水平和CD8+T细胞中毒性分子mRNA水平, 流式细胞术检测CD8+T细胞中免疫检查点分子水平。组间比较采用t检验或Mann-Whitney检验。结果重组EBNA2刺激后外周血单个核细胞增殖、CD4+T细胞和CD8+T细胞比例在EBV感染无肝脏损伤组和EBV感染致肝脏损伤组之间的差异无统计学意义(P > 0.05)。重组EBNA2刺激后CD4+T细胞分泌相关细胞因子比例和转录因子mRNA水平在EBV感染无肝脏损伤组和EBV感染致肝脏损伤组之间的差异无统计学意义(P...     

Parallel decline of CD8＋CD38＋ lymphocytes and viremia in treated hepatitis B patients

AIM:To assess the peripheral T lymphocyte subsets in chronic hepatitis B virus(HBV) infection,and their dynamics in response to adefovir dipivoxil monotherapy.METHODS:Proportions and absolute counts of peripheral natural killer cells,B cells,CD8+,CD4+,CD8+ CD38+,CD8+CD28+ and CD4+CD28+ T cells were determined using three-color flow cytometry in chronic hepatitis B patients(n = 35),HBV carriers(n = 25) and healthy controls(n = 35).Adefovir dipivoxil was initiated in 17 chronic hepatitis B patients who were r...     

Expansions of CD8+CD28- and CD8+TcRVbeta5.2+ T cells in peripheral blood of heavy alcohol drinkers

BACKGROUND: Despite heavy alcohol consumption, only a low percentage of heavy drinkers develop liver disease. Imbalances in T-cell subsets and iron metabolism parameters are common findings in heavy drinkers, yet the possible role played by discrete T-lymphocyte subsets under heavy alcohol consumption remains unclear. METHODS: To gain new insights into the possible role played by T lymphocytes during alcohol consumption, characterization of CD28 expression and TcR repertoire in peripheral blood CD4+ and CD8+ T cells by two and three-color flow cytometry was performed. A group of heavy alcohol drinkers (AHD, n = 71) and a group of age-matched controls (n = 81), both HLA-phenotyped and HFE-genotyped, constituted the groups under study. RESULTS: Marked expansions of CD28- T cells within the CD8+ but not the CD4+ T-cell pool were observed in AHD compared with controls. These CD8+CD28- expansions were paralleled by expansions of CD8+ T cells bearing specific TcR Valpha/beta chains, namely VP5.2. Moreover, AHD, but not controls, carrying the H63D mutation in the HFE gene showed significantly higher percentages of CD28- T cells within the CD8+ T-cell pool than AHD carrying the normal HFE gene. Finally, high numbers of CD8+CD28- T cells in AHD were associated with lower levels of the liver-related enzymes ALT and GGT. CONCLUSIONS: This study showed that under active ethanol consumption, expansions of discrete CD8+ T-cell subsets occur within the CD8+ T-cell pool, that molecules of the MHC-class I locus seem to influence the extent of the expansions, and that high numbers of CD8+CD28- T cells are associated with low levels of liver enzymes in AHD.     

强直性脊柱炎患者外周血T淋巴细胞亚群和T淋巴细胞上CD154表达的变化和意义

目的 探讨强直性脊柱炎(AS)患者外周血T淋巴细胞亚群分布和T淋巴细胞上共刺激分子CD154的表达及依那西普治疗对其的影响.方法 用流式细胞仪检测66例AS患者(其中活动期39例,非活动期27例;按临床特征分为外周关节和中轴均受累者35例和单独中轴受累31例)、30例类风湿关节炎(RA)患者及30名健康志愿者外周血T淋巴细胞亚群分布和CD154在CD3+T淋巴细胞上的表达.此外,观察39例AS活动期患者在随机双盲依那西普和安慰剂对照试验中,用药前后CD154的变化.结果 ①AS和RA患者CD4+T淋巴细胞均较健康志愿者高(P＜0.05),而CD8+T淋巴细胞较健康志愿者低(P＜0.05);AS患者外周血T细胞上CD154的表达较健康志愿者和RA患者都明显升高(P＜0.05);②活动期或外周关节受累AS患者T细胞CD154的表达分别较稳定期或单独中轴受累AS患者明显升高(P＜0.05);且CD154的表达与关节压痛数、关节肿胀数呈正相关(P＜0.05);③在依那西普试验的第6周,依那西普组AS患者(19例)T淋巴细胞CD154表达较安慰剂组(20例)明显下降(P＜0.05),与健康志愿者差异无统计学意义(P＞0.05).结论 AS患者外周血存在T淋巴细胞亚群紊乱和T淋巴细胞上共刺激分子CD154异常表达,可作为临床评价AS病情活动性和依那西普疗效的生物指标之一.     

Upregulated expression of VLA proteins and CD29 in peripheral blood lymphocytes of chronic alcoholics without ethanol-related diseases

To analyze adhesion molecule expression on peripheral blood mononuclear cells (PBMCs) and on different lymphocyte subpopulations (CD2+, CD8+, CD19+, and CD56+ subsets) in chronic alcoholism, 30 well-nourished chronic alcoholics without ethanol-related diseases and 30 matched controls were included in the study. Adhesion molecules that mediate adhesion to other cells and to extracellular matrix proteins, and whose cellular expression is modified during lymphocyte activation, were selected for study. A detailed clinical evaluation, laboratory analysis, nutritional assessment, and study of adhesion molecule expression was performed. A significant higher expression of CD29 (beta1-integrin) (p = 0.001), VLA-3 (p = 0.002), VLA-4 (p = 0.03), and VLA-5 (p = 0.001) were observed on PBMCs of chronic alcoholics, compared with control subjects, whereas no changes were observed in CD18 (beta2-integrin) and CD50 (ICAM-3) expression. The upregulation of CD29 and VLA proteins only affected T lymphocytes (CD2+/CD8+/CD4+ cells). These data confirm that T cells of chronic alcoholics are basally activated and that changes in adhesion molecule expression on PBMCs may be responsible of disturbances of adhesion processes in chronic alcoholics without ethanol-related diseases.     

Expression of CD44 variant isoforms CD44v3 and CD44v6 is increased on T cells from patients with systemic lupus erythematosus and is correlated with disease activity

Objective

To quantify the expression of CD44 and variant isoforms CD44v3 and CD44v6 on T cells from patients with systemic lupus erythematosus (SLE), and to assess correlations of the level of expression of these molecules with disease manifestations.

Methods

Information on clinical and demographic characteristics was collected, and blood samples were obtained from 72 patients with SLE and 32 healthy control subjects matched to the patients by sex, race, and age. Expression of CD44 and variants CD44v3 and v6 on T cell subsets was determined by flow cytometry, and Pearson's correlations of their expression levels with clinical variables, SLE Disease Activity Index (SLEDAI) scores, and presence of lupus nephritis were determined. Wilcoxon's rank sum tests and conditional multivariable regression analyses were applied to identify differences in the expression of CD44 between patients with SLE and healthy controls.

Results

Expression of CD44 was higher on CD4+ and CD8+ T cells from SLE patients compared with controls (P ≤ 0.03). Expression of CD44v3 and CD44v6 was also higher on total T cells and CD4+ and CD8+ T cells from SLE patients compared with controls (P ≤ 0.03). Cell surface levels of CD44v3 on total T cells, CD4+ T cells, and CD8+ T cells as well as cell surface expression of CD44v6 on total T cells and CD4+ T cells were correlated with the SLEDAI score (P < 0.05). The presence of lupus nephritis was associated with the expression of CD44v6 on total T cells, CD4+ T cells, and CD4−CD8− T cells (P < 0.05). Positivity for anti–double‐stranded DNA antibodies was associated with the expression levels of CD44v6 on T cells (P < 0.05).

Conclusion

These results indicate that expression levels of CD44v3 and CD44v6 on T cells may represent useful biomarkers of SLE activity.

     

IL-7 concentration is increased in nonagenarians but is not associated with markers of T cell immunosenescence

Interleukin-7 is a homeostatic cytokine that contributes to the maintenance of the T cell pool. It also has proinflammatory effects and is involved in the pathogenesis of autoimmune diseases. Due to its homeostatic effects, IL-7 has been proposed as a potential rejuvenation factor for the elderly immune system. We analyzed the correlation of plasma IL-7 concentrations and the proportions of different T cell populations in nonagenarians (n=163) participating in the Vitality 90+ study. Young individuals (n=35, aged 19-30years) were used as controls. The numbers of CD3+, CD14+, CD4+ and CD8+ cells and the expression of the CD28 costimulatory molecule on CD4+ and CD8+ lymphocyte subsets were analyzed using flow cytometry. The plasma IL-7 levels were significantly higher in the nonagenarians compared to the controls (7.86 vs. 5.74pg/ml, p=0.004). In the nonagenarians, plasma IL-7 levels correlated inversely with the proportion of CD3+ T cells and directly with the proportion of CD14+ monocytes and plasma C-reactive protein. No correlation was observed between plasma IL-7 levels and the proportions of CD4+CD28- or CD8+CD28- subsets. These results suggest that the IL-7 levels in nonagenarians do not have an inhibitory effect on the development of immunosenescence; rather they are associated with increased inflammation.     

Characterization of the expanded T-cell populations in patients with Wegener's granulomatosis

OBJECTIVES: Wegener's granulomatosis (WG) is a chronic inflammatory disease characterized by granulomatosis inflammation, systemic vasculitis and glomerulonephritis. In patients, the peripheral T cells are characterized by mono/oligoclonal CD4+/CD8+ T-cell AV/BV receptor expansions, with aberrant expression of activation markers. This study was designed to characterize the phenotypic differences between the expanded and nonexpanded T-cell populations. Expression of markers for activation, costimulation and adhesion molecules was examined. As earlier studies have shown aberrant expression of CD28/CD152, we also analysed the expression of another costimulatory system, the tumour necrotic factor receptor (TNFR) superfamily proteins. DESIGN: Fluorocrome-conjugated monoclonal antibodies and flow cytometry was used to analyse the expression of the different markers on the surface of the expanded and nonexpanded subsets of T cells. SETTING: The Karolinska Hospital and Karolinska Institutet in Stockholm, Sweden. SUBJECTS: Nine patients with WG (six men and three women) had 16 TCRAV/BV CD4+/CD8+ expanded populations that were characterized. RESULTS: The expanded TCRA/BV CD4+ and CD8+ cells had lower percentages of cells expressing CD28 and higher of those expressing CD152 (CTLA-4). The expanded CD4+ population had more cells expressing HLA-DR, CD57 and CCR5 (CD195), whilst the expression of CD25 was present on fewer of the expanded cells. The expanded CD8+ population contained more cells expressing CD137 (4-1BB), CD137 (4-1BBL), CD30 (Ki-1), CD40 and CD134 (OX40). CONCLUSIONS: There were marked differences in the phenotypes of expanded and nonexpanded T-cell populations.