老年人外周血T细胞亚群CD28表达水平的改变

21世纪人口老龄化对人类社会是一个重大的挑战。近年来,越来越多的研究结果表明免疫反应性与老年个体的健康状态之间有着密切的关系。免疫系统的衰老是老年个体感染性疾病、癌症、自身免疫性疾病发生率、病死率增高的主要原因,此外,免疫衰老与个体寿命间的联系也得到了大量证据     

再生障碍性贫血患者外周血T淋巴细胞亚群及HLA-DR抗原的变化及临床意义

目的：探讨再生障碍性贫血（再障）患者治疗前、后外周血T淋巴细胞亚群及HI。A-DR抗原的变化及其临床意义。方法：采用流式细胞术检测28例再障患者治疗前、后外周血T淋巴细胞亚群及HLA-DR抗原的变化，并与健康对照组进行比较。结果：①再障患者治疗前CD4^＋、CD4^＋／CD8^＋明显低于健康对照组，CD8^＋、HLA—DR^＋则明显升高；重型再障组与普通型再障组比较，CD8^＋、HLA-DR^＋明显升高，CD3^＋、CD4^＋、CD4^＋／CD8^＋无统计学意义。②再障患者治疗后与治疗前相比，CD4^＋、CD4^＋／CD8^＋明显升高，CD8^＋、HLA-DR^＋明显降低。③再障患者治疗有效者与无效者比较，CD4^＋、CD4^＋／CD8^＋明显升高，CD8^＋、HLA-DR^＋明显降低。结论：再障患者存在T淋巴细胞亚群的失调及T淋巴细胞的异常活化，检测外周血T淋巴细胞亚群及HLA-DR抗原有助于再障患者的病情监测、疗效评价及预后判断。     

病毒性心肌炎患者外周血T淋巴细胞亚群及B淋巴细胞的研究

目的: 测定不同时期病毒性心肌炎(VMC)患者T淋巴细胞亚群及B淋巴细胞的数量,并同时检测患者血清心肌损害标志物心肌肌钙蛋白I(cTnI)的水平,探讨其在VMC发病机制中的作用。方法: 采用流式细胞术检测不同时期VMC患者(n=126)外周血CD4+CD45RA+和CD4+CD45RO+以及CD19+的表达,并设正常人群作为对照组(n=50)。结果: 急性期（<3个月）患者CD4+CD45RA+和CD4+CD45RO+以及CD19+与对照组相比差异显著。<1个月以CD4+CD45RA+和CD4+CD45RO+为著（P<0.01;P<0.01）。病程<7 d的患者与对照组相比,CD4+CD45RA+值的降低具有显著性(P<0.01);CD4+CD45RO+值下降明显(P<0.01)。而病程7~14 d、14~21 d及21~30 d的患者CD4+CD45RA+值、CD4+CD45RO+值及CD19+值与对照组相比差异均有统计学意义,CD4+CD45RO+值恢复较快。CD4+CD45RA+/CD4+CD45RO+在发病14 d内的患者与对照组差异无统计学意义;而14~30 d的患者与对照组相比差异有显著性。发病1~3个月患者以CD19+为著（P<0.01）。急性期CD4+CD45RA+/CD4+CD45RO+细胞的比例失衡。恢复期（3~12个月）患者CD4+CD45RA+和CD4+CD45RO+以及CD19+、CD4+CD45RA+/CD4+CD45RO+细胞的比例与对照组相比差异无显著性。结论: VMC患者感染初期（<1个月）以T淋巴细胞亚群异常为主,感染后期（1~3个月）以B淋巴细胞异常较为显著,以上两时期均存在CD4+CD45RA+/ CD4+CD45RO+细胞比例失衡。恢复期（3~12个月）T淋巴细胞及B淋巴细胞逐渐恢复至正常。     

不同心功能患者T淋巴细胞亚群的变化

目的研究不同心功能分级患者外周血T淋巴细胞亚群的变化。方法健康体检者39例为对照组,器质性心脏病患者84例,根据心功能分级标准(NYHA),心功能Ⅰ级组33例,Ⅱ级组20例、Ⅲ级组16例、Ⅳ级组15例。采用流式细胞仪测定外周血T淋巴细胞亚群CD3、CD4、CD8,计算CD4/CD8比值,超声心动图测定左心室射血分数(LVEF)。结果T淋巴细胞亚群CD3水平在对照组和心功能Ⅰ、Ⅱ、Ⅲ、Ⅳ级组之间差异无显著性;CD4水平从心功能Ⅰ级至Ⅳ级组逐渐降低,对照组及心功能Ⅰ级组与心功能Ⅲ级组、Ⅳ级组比较,差异均有显著性(P<0.01),心功能Ⅱ级组与Ⅳ级组比较差异也有显著性(P<0.05);CD8水平在心功能Ⅳ级组明显升高;CD4/CD8与CD4变化相一致,随心功能减退而降低。结论心力衰竭患者存在细胞免疫功能异常,随心功能减退辅助性T淋巴细胞数量减少,提示调节细胞免疫治疗可能会使心力衰竭患者获益。     

肝炎肝硬化及原发性肝癌患者外周血Ｔ淋巴细胞亚群分析

为了探讨慢性肝病合并原发性肝细胞癌患者细胞免疫功能紊乱的发病机理,我们运用流式细胞技术对39例肝炎肝硬化及35例慢性病毒性肝炎合并原发性肝癌患者外周Ｔ血淋巴细胞亚群进行了检测。现将结果分析如下。对象与方法1.对象:74例患者均为1997年1月～1997年6月我院住院病人,分为肝炎肝硬化组(39例)和原发性肝癌组(35例)。肝炎肝硬化的诊断标准符合1995年全国传染病与寄生虫病学术会议修订的《病毒性肝炎防治方案》(北京)标准,男性30例,女性9例,年龄为31～60岁,平均年龄(42.4±11.6)岁。39例肝炎肝硬化患者乙肝病毒标志物均为阳性,其中5例合并…     

93例传染性非典型肺炎患者外周血T淋巴细胞亚群变化及临床意义

目的　了解传染性非典型肺炎 (世界卫生组织又称严重急性呼吸综合征 ,SARS)患者外周血T淋巴细胞亚群的变化。方法　采用流式细胞仪对 93例临床确诊的SARS患者、5 0例获得性免疫缺陷综合征 (AIDS)患者及 6 4例健康体检者外周血T淋巴细胞亚群进行检测。 93例患者中 ,男4 0例、女 5 3例 ;年龄 17～ 88岁 ,平均 4 4岁 ;重型 35例、普通型 5 8例。结果　健康体检者外周血CD+ 3 、CD+ 4 、CD+ 8分别为 (15 2 7± 4 70 )、(787± 2 5 7)、(6 33± 2 80 )个 / μl;93例急性期SARS患者分别为(72 2± 5 33)、(438± 35 3)、(30 7± 2 17)个 / μl,均有不同程度的下降 (P值均 <0 .0 1) ,重症病例下降尤其明显 ,5例死亡患者外周血CD+ 4 均低于 2 0 0个 / μl;SARS患者恢复期CD+ 3 、CD+ 4 、CD+ 8多数恢复正常。而AIDS患者以CD+ 4 降低为主 ,为 (2 96± 2 98)个 / μl;且CD+ 8升高 ,为 (818± 5 6 6 )个 / μl。 结论SARS患者有明显的细胞免疫损伤。     

肺结核患者T淋巴细胞亚群的检测及临床意义

目的探讨肺结核患者T淋巴细胞亚群的变化及临床意义。方法用流式细胞术检测179名肺结核患者静脉全血CD3、CD4、CD8T淋巴细胞绝对计数和CD4/CD8比值,与110名健康人进行对照,观察肺结核患者与健康人之间,各型肺结核之间及菌阳与菌阴、初治与复治之间的差别。结果肺结核患者CD3、CD4T淋巴细胞绝对计数、CD4/CD8比值明显下降,CD8T淋巴细胞绝对计数增高,与健康人对比差异有统计学意义(P<0.01),各型结核之间、痰菌阳性与阴性结核之间、初治与复治结核之间也有一定的差异,但无统计学意义(P>0.05)。结论T淋巴细胞亚群可作为判断肺结核患者免疫功能的指标,这对于了解肺结核患者的免疫状态、指导治疗、考核疗效有一定的临床意义。     

乙型肝炎患者外周血T淋巴细胞亚群的检测

乙型病毒性肝炎(乙肝)的肝细胞损害和炎症反应是免疫细胞作用于肝细胞的结果,以T淋巴细胞破坏已感染乙型肝炎病毒(HBV)的肝细胞的作用最为重要.我们对乙肝患者外周血T淋巴细胞亚群和e抗原系统之间的关系进行了探讨.     

小细胞肺癌患者外周血淋巴细胞亚群分析

目的观察小细胞肺癌(SCLC)患者外周血中T淋巴细胞亚群的水平,探讨SCLC免疫变化及其意义。方法采用流式细胞技术,根据白细胞表面抗原分化群(CD)检测33例SCLC患者外周血中的CD+3、CD+4、CD+8、CD+19及CD+16CD+56、活化T细胞比例,以30例健康人做对照,结合临床病理及随访资料,比较其差异。结果 SCLC患者外周血CD+3、CD+19细胞较对照组低;活化T细胞两组间无明显统计学意义。结论SCLC患者机体的细胞免疫异常,与病情发展、临床分期密切相关。检测T淋巴细胞亚群有助于观察患者的免疫状态。     

乙型肝炎患者外周血T淋巴细胞亚群的变化

目的分析乙型肝炎病毒(HBV)感染者外周血淋巴细胞亚群的变化,以了解细胞免疫在乙型肝炎发病中的作用。方法使用流式细胞仪检测100例乙型肝炎患者和20例正常人外周血CD3+、CD4+和CD8+T淋巴细胞亚群百分比。结果 15例急性肝炎患者T淋巴细胞亚群与正常对照组比,差异无统计学意义(P>0.05),而43例慢性肝炎、26例乙型肝炎肝硬化和16例慢性肝衰竭患者CD3+淋巴细胞百分率分别为56.15±8.94%、48.85±9.01%和55.86±9.03%,均显著低于正常对照组的66.35±8.93%(P<0.05);慢性肝炎、乙型肝炎肝硬化和慢性肝衰竭患者CD4+淋巴细胞百分率分别为30.46±7.51%、26.71±7.03%和29.03±7.64%,,均显著低于正常对照组的35.72±7.52%(P<0.05);慢性肝炎、乙型肝炎肝硬化和慢性肝衰竭患者CD8+淋巴细胞百分率分别为21.93±5.06%、18.71±5.53%和21.15±5.62%,均显著低于正常对照组的24.58±4.92%(P<0.05);慢性肝炎、乙型肝炎肝硬化和慢性肝衰竭患者CD4+/CD8+比值分别为1.34±0.60、1.35±0.53和1.34±0.58,均低于正常对照组的1.58±0.47(P<0.05)。结论 HBV感染者体内存在T细胞亚群失衡和细胞免疫功能紊乱,提示细胞免疫参与了乙型肝炎的发病和疾病进展。     

老年非小细胞肺癌外周血CD25和HLA-DR^＋表达的临床研究

目的观察老年非小细胞肺癌患者外周血T淋巴细胞活化抗原CD3^＋／CD25和CD3^＋／HLA-DR的表达，进一步探讨肺癌的免疫功能。方法应用流式细胞双色免疫荧光标记技术对45例老年肺癌患者外周血T淋巴细胞活化抗原CD3^＋／CD25和CD3^＋／HLA—DR表达进行荧光免疫检测，并与中青年肺癌组、正常老年对照组和老年良性病变组，进行对比研究。结果45例老年肺癌患者外周血T淋巴细胞中CD3^＋／CD25^＋和CD3^＋／HLA-DR表达明显低于正常对照组和良性病变组（P〈0．01）。老年组和中青年组之间其表达水平比较也有显著性差异（P〈0．05或〈0．01）。正常老年对照组和老年良性病变组之间无显著性差异（P〉0．05）。Ⅲ期、Ⅳ期和Ⅰ期、Ⅱ期之间CD3^＋／CD25^＋和CD3＋／HLA-DR表达比较，差异有显著性（P〈0．01）。结论检测外周血T淋巴细胞活化抗原CD3^＋／CD25^＋和CD3^＋／HLA-DR^＋的表达水平对了解老年肺癌患者的免疫状态和预后评估具有十分重要的意义。     

The naive T-lymphocyte compartment is well preserved in patients with chronic myelogenous leukaemia in chronic phase

In chronic myelogenous leukaemia (CML), clonal change occurs in all myeloid and B-cell lineages, but very rarely T-cell lineages. A detailed three-colour cytometric analysis of peripheral lymphocytes was performed in 22 patients with chronic-phase CML (CP-CML). CD45 gating analysis was used to discriminate between lymphocytes and basophils. The peripheral lymphocyte pool was comprised of a significant proportion of naive CD4 cells, defined by a CD4+45RA+ phenotype [47.0 +/- 19.6% (mean +/- SD) of the total CD4+ cells], and naive CD8 cells, defined by a CD8+CD45RA+CD28+ phenotype (35.1 +/- 19.7% of total CD8+ cells), even in patients with long disease duration. The percentage of CD8 naive T cells showed inverse correlation with age, whereas no correlation was observed with disease duration. Possible explanations for the preservation of naive lymphocytes include (1) that the naive T cells differentiated from co-existing normal stem cells or (2) that long-lived naive T cells persisted from the CML onset and expanded peripherally (thymus independent). Either mechanism or a combination of both mechanisms might contribute to maintaining the naive compartment size.     

支气管肺泡灌洗液T淋巴细胞亚群和细胞DNA含量分析对肺癌的诊断价值

目的　探讨支气管肺泡灌洗液T淋巴细胞亚群和细胞DNA含量分析对肺癌的诊断价值。方法　用流式细胞仪对 3 0例肺癌和 12例肺良性病变患者的支气管肺泡灌洗液 (BALF)作T淋巴细胞亚群和细胞DNA含量分析 ,并与纤维支气管镜活检和刷检比较。结果　中央型肺癌组BALF中CD3 + (11.473± 3 .677) %、CD4+ (3 .660± 1.60 5 ) %、CD8+ (2 .3 49± 0 .880 ) %和CD4+ /CD8+ (1.411± 0 .3 0 9) ,周围型肺癌组BALF中CD3 + (12 .2 73± 4.42 5 ) %、CD4+ (2 .897±0 .695 ) %、CD8+ (2 .2 5 9± 0 .5 91) %和CD4+ /CD8+ (1.3 0 7± 0 .2 45 ) ;肺良性病变组BALFCD3 +(2 5 .0 78± 7.13 8) %、CD4+ (13 .2 44± 4.15 9) %、CD8+ (7.63 1± 3 .713 ) %和CD4+ /CD8+ (2 .2 78±0 .619) ,前两组显著低于后组 (肺良性病变组 ) (P <0 .0 0 1)。以异常二倍体为阳性标准 ,诊断肺癌的敏感性为 83 % ,特异性为 92 % ;中央型肺癌异常二倍体阳性率为 89% ,与活检 (95 % )和刷检(68% )比较 ,差异无显著性 (P >0 .5和P >0 .1) ;周围型肺癌异常二倍体阳性率为 82 % ,显著高于活检 (2 7% )和刷检 (2 7% )阳性率 (P均 <0 .0 5 )。结论　肺癌患者免疫功能低下 ,是肿瘤发生发展的原因之一 ,支气管肺泡灌洗液细胞DNA含量分析是诊断肺癌 (尤     

Analysis of T-lymphocyte subpopulations in inflammatory bowel diseases by three-color flow cytometry

In order to better define changes in the relative proportion of peripheral blood T-lymphocyte subpopulations in patients with inflammatory diseases of the bowel, we performed simultaneous three-color fluorescence-activated cytometric (FACS) analysis using fluorophore-conjugated monoclonal antibodies with specificity for CD4, CD8, Leu 8, and CD45RA on 22 normal control subjects, 28 patients with Crohn's disease (CD), 15 patients with ulcerative colitis (UC), and 11 patients with intestinal inflammation secondary to etiologies other than inflammatory bowel disease (NIBD). This staining combination allowed enumeration of distinct T-cell subpopulations as follows: virgin CD4⁺, recall antigen helper T cells, nonspecific B-cell helper T cell, virgin CD8⁺, cytotoxic effector and suppressor effector and recall antigen cytotoxic T cells based on a synthesis of published functional analyses. No differences in the proportion of CD4⁺ or CD8⁺ cells or in the CD4⁺/CD8⁺ ratios were evident when UC and NIBD patients were compared to normal subjects. A significant reduction in the proportion of CD4⁺ cells and an increase in CD8⁺ cells was observed, however, in the CD group. When two-color analysis was performed, several significant differences in the proportions of circulating lymphocytes were seen. Specifically, these included significant increases in the number of CD4⁺, Leu 8^– (P<0.01) cells in all disease groups and an increase in CD4⁺, CD45RA⁺ cells in the NIBD group. Conversely, significant decreases in the proportions of CD8⁺, Leu 8⁺ (P<0.01) cells were evident in the Crohn's disease group. Three-color FACS analysis revealed significant differences in the relative proportions of the defined T-cell subpopulations enumerated in the various groups as compared with the normal controls. These included a decrease in the proportions of Leu 8⁺, CD45RA⁺, (virgin) CD8⁺ T cells (P<0.05) and Leu 8^–, CD45RA⁺, (putative recall antigen helper) CD4⁺ T cells (P<0.01) in all patient groups as compared with normal controls. Conversely, an increase in the proportions of Leu 8^–, CD45RA⁺, (putative suppressor effector) CD8⁺ T cells (P<0.01), and Leu 8^–, CD45RA⁺, (function unknown) CD4⁺ T cells (P<0.05) was seen in all patient groups as compared with normal controls. CD patients but not UC or NIBD patients demonstrated a significant increase in the proportion of Leu 8^–, CD45RA⁺, (putative cytotoxic effector) CD8⁺ T cells (P<0.01). An increase in the ratio of Leu 8^–, CD45RA⁺, CD8⁺ (suppressor effector)/Leu 8⁺, CD45RA^–, CD8⁺ (putative cytotoxic effector) T cells was observed in all of the patient groups, but was most accentuated in those with Crohn's disease. Significant decreases in the ratios of Leu 8^–, CD45RA^–, CD4⁺ (putative nonspecific B cell helper)/Leu 8⁺, CD45RA^–, CD4⁺ (recall antigen helper) T cells and Leu 8⁺, CD45RA^–, CD4⁺ (recall antigen helper)/Leu 8^–, CD45RA⁺, CD4⁺ (function unknown) T cells were observed in all of the disease groups studied as compared with normal controls. These results suggest that the proportions of certain peripheral blood T-cell subpopulations are significantly altered in gastrointestinal inflammatory states. Further analysis of these T-lymphocyte subpopulations in the blood and tissues might provide valuable insights into immunological aberrations in inflammatory bowel diseases and might be of value in distinguishing among inflammatory diseases of the intestine.     

胃癌病人血清SIL-2R和外周血CD4/CD8的相关研究

分别应用双抗体夹心法和流式细胞术测定52例胃癌病人的血清可溶性白细胞介素2受体(Soluble inter-leukin2 receptor,SLI-2R)和外周血CD_4/CD_8,结果胃癌病人SIL-2R明显升高,CD_4/CD_8明显降低(均P<0.01),且临床分期越高越明显,两者之间存在明显的负相关关系(r=-0.87),提示胃癌病人细胞免疫功能抑制。     

戊型肝炎患者T淋巴细胞亚群的变化

目的探讨戊型肝炎患者外周血T淋巴细胞亚群的变化及其免疫学意义。方法应用流式细胞仪检测136例急性戊型肝炎患者及13例正常人群的外周血T细胞亚群。结果戊型肝炎患者与正常人比较,T淋巴细胞亚群变化无统计学差异（P〉0.05）。单纯戊型肝炎病毒感染组的CD3＋细胞总数、CD4＋和CD8＋细胞绝对数均高于重叠乙型肝炎病毒感染组（P均〈0.05）。结论戊型肝炎病毒感染患者存在宿主细胞免疫功能的改变,这可能与急性戊型肝炎肝细胞损伤和清除病毒有关。     

Changes in the expression of surface receptors on lymphocyte subsets in the elderly: quantitative flow cytometric analysis

The immunophenotype of circulating lymphocytes, including the intensity expression of surface receptors, changes with ageing. Until now, no results of systematic studies on age-dependent changes with respect to the expression of the major lymphocyte surface receptors in healthy elderly subjects have been reported. In order to identify age-related changes in both representation and immunophenotype of lymphocyte populations, we investigated, by means of triple-color whole-blood immunostaining and quantitative flow cytometry, the percent values and the absolute numbers, as well as the levels of surface antigen expression or antigen molecules per cell (ABC values x 10(3)), of different peripheral blood lymphocyte subsets from 23 healthy elderly subjects and 13 young donors. Naive (CD45RA+CD3+) T cells, total B cells, and CD5+ B lymphocytes are decreased (22%, 6%, 0.8% vs. 30%, 12%, 1.4%, respectively), whereas activated (HLA-DR+CD3+) and memory (CD45RO+CD3+) T cells, CD3+CD7- T lymphocytes, and lymphocytes expressing the NK marker CD56 are expanded in the elderly (2%, 53%, 13%, 6% vs. 0.8%, 45%, 8%, 8%, respectively). Moreover, T lymphocytes from elderly individuals express lower CD3 (61 +/- 10) compared to young (69 +/- 10). Considering the different T-cell populations, CD3 antigen is respectively decreased on CD45RO+ T cells (55 +/- 14 vs. 66 +/- 14) and up-regulated on CD56+ T lymphocytes (62 +/- 21 vs. 45 +/- 20). Increased CD8 expression characterizes CD3+CD7- lymphocytes (70 +/- 34 vs. 44 +/- 17) while HLA-DR on activated T cells is lower in old (39 +/- 7) than young (46 +/- 9) donors. CD7 is down-regulated both in T (22 +/- 3 vs. 28 +/- 3) and NK (48 +/- 18 vs. 71 +/- 18) cells, whereas CD2 expression, unchanged on NK cells, is up-regulated on T lymphocytes (54 +/- 10 vs. 41 +/- 8). Age-related changes in B-cell antigen expressions were also found: CD20 is increased (124 +/- 23 vs. 105 +/- 16) whereas, despite the unchanged CD5 expression of T cells, CD5 intensity on the B-cell subset co-expressing this antigen is higher in old (49 +/- 37) than in young (22 +/- 4) people. The observed changes in the expression of functionally important cellular receptors can contribute to the remodeling of immune function characteristic of the elderly. Moreover, since quantitative flow cytometry is becoming widely employed in clinical practice, our results also contribute to the assessment of specific age-dependent antigen expression changes to be considered for diagnostic approaches in the elderly.     

Normal human blood density gradient lymphocyte subset analysis: I. An interlaboratory flow cytometric comparison of 85 normal adults

An interlaboratory flow cytometric comparison of several commercially available human lymphocyte subset reagents was undertaken in three different laboratories. Fresh Hypaque-Ficoll purified blood mononuclear cells were stained at 4 degrees C or 22 degrees C. Direct or indirect surface immunofluorescence was carried out at all sites using an EPICS V flow cytometer. Fullbright, 10-micron fluorescent polystyrene microspheres were used for optical alignment and standardization. A log integral fluorescent histogram gated on forward and right angle scatter was collected on 1-2 X 10(4) cells for each reagent and the proportion, of positive cell determined for each reagent. With the exception of one reagent, anti-B1, which showed an approximately twofold variation, all three laboratories showed remarkable agreement. Thus there was no significant difference noted for the following reagents: OKT4, CCT4, Leu 3a, Leu 2a, OKT8, or CCT8. We attribute these findings to the availability of quality reagents, precision instrumentation, and a standard lymphocyte preparation.     

肝硬化患者外周血及腹水T淋巴细胞亚群的变化及意义

目的研究肝硬化患者外周血及腹水T淋巴细胞亚群的变化及意义。方法流式细胞仪测定31例肝炎后肝硬化患者外周血及腹水T淋巴细胞亚群，同时检测15例正常人的外周血T淋巴细胞亚群。结果肝硬化患者外周血T淋巴细胞（CD3^＋）、T辅助／诱导细胞亚群（CD4^＋）、T辅助淋巴细胞／T抑制淋巴细胞亚群（CD4^＋／CD8^＋）较正常对照明显降低（p＜0．01），腹水CD3^＋、CIM^＋、CD8^＋、CD4^＋／CD8^＋较外周血显著降低（P＜0．001）。结论肝硬化患者外周血及腹水T淋巴细胞亚群存在明显异常，表现为全身及腹膜腔局部的免疫力低下。