NK相关抗原在HIV/AIDS患者CD8+T淋巴细胞上的表达

目的　探讨NK相关抗原CD5 6、CD16在HIV AIDS患者CD8 T淋巴细胞上的表达。方法　取外周血细胞 ,用标记荧光的抗体进行染色 ,以CD8强阳 淋巴细胞为门 ,用流式细胞仪分析CD8 T淋巴细胞上CD5 6、CD16的表达。结果　HIV AIDS患者CD8 T淋巴细胞表达CD5 6 、CD5 6 CD16 - 、CD5 6 CD16 均明显低于HIV抗体阴性健康对照组 (P <0 .0 5 ) ;经高效抗逆转录病毒疗法 (HAART)治疗后CD8 T淋巴细胞表达的CD5 6 、CD5 6 CD16 - 、CD5 6 CD16 呈逐渐升高趋势。HIV AIDS患者表达CD5 6 CD16 - 的CD8 T淋巴细胞亚群绝对数与CD4 T淋巴细胞绝对数呈正相关 ,r=0 .393,P <0 .0 5 ;表达CD5 6 - CD16 的CD8 T淋巴细胞百分数与CD4 T细胞绝对数呈负相关 ,r=- 0 .32 4 ,P <0 .0 5。结论　表达CD5 6的CD8 T淋巴细胞在HIV AIDS患者中明显缺失 ,HAART治疗可恢复缺失。CD8 T淋巴细胞上CD5 6的表达是HIV感染中值得关注的重要指标之一 ,对评价抗病毒疗效具有指导意义。     

医源性HIV感染者CD8+细胞非细胞毒性HIV抑制作用的研究

目的 观察医源性HIV感染者CD8+细胞非细胞毒性HIV抑制反应(CNAR),并比较CNAR与CD4细胞计数的关系.方法 免疫磁珠法分离HIV感染者的CD8+细胞,按2:1,1:1,0.5:1和0.25:1的比例,与体外急性感染的CD4+细胞混合培养,测定培养物上清中逆转录酶活性,与阴性对照比较计算病毒抑制率.结果 CNAR活性达到80%病毒抑制率时,CD4<300个/μl组的平均CD8与CD4比例为2.4:1,CD4>300个/μl组的平均CD8与CD4比例为1.3:1,两组间比较差异有统计学意义(P<0.05).结论 HIV感染者的CNAR活性与其CIM细胞计数相关,CD4>300的个体较CD4<300的个体有着更显著的抑制HIV复制的能力.     

γδT、CD4 +CD25 +T细胞在HIV感染者/AIDS患者外周血的表达及其相关性研究

目的:了解γδT细胞、CD4 +CD25 +调节性T细胞(Treg)在HIV感染者/AIDS患者外周血中的表达水平,探讨γδT细胞、CD4 +CD25 + Treg在HIV感染/AIDS病情进展中的相关性以及可能的作用机制。 方法:采用免疫荧光单克隆抗体标记技术...     

补体调节蛋白CD59在HIV感染者外周血T细胞上的表达研究

目的 观察补体调节蛋白CD59在HIV感染者外周血T细胞上的表达情况,并进一步探讨高效抗逆转录病毒治疗（Highly Active Antiretroviral Therapy,HARRT）前后CD59在外周血T细胞上的表达变化及其意义.方法 收集48例确诊HIV感染者外周血,根据是否HARRT分为未治疗组（23例）及治疗组（25例）,并收集14例健康志愿者外周血,全血细胞表面染色后使用BD FACSCanto 流式仪检测CD59在外周血T细胞上的表达情况,并进一步检测CD59在CD45RO＋记忆型CD4T细胞表面的表达情况;同时用定量PCR技术检测HIV感染者的血浆病毒载量及CD4细胞绝对计数,将结果进行统计学分析并比较各组间差异.结果 与健康对照者比较,CD59在HIV感染者CD4＋T细胞上的表达水平明显增高（P＜0.05）,其中以在CD45RO＋记忆型CD4T细胞上增高为主（P＜0.05）;治疗组CD4＋T细胞上CD59的表达较未治疗组明显下降（P＜0.05）,但与健康对照组比较其表达仍处于较高水平（P＜0.05）,进一步分析发现CD59的这种变化主要发生在CD45RO＋（记忆型）CD4＋T细胞上;CD59在CD4＋T细胞上的表达变化与病毒载量及外周血CD4＋T细胞绝对计数均有一定的相关性（R2=0.2181,P=0.0247;R2=0.1586,P=0.0486）.结论 HIV感染可引起补体调节蛋白CD59在CD4＋T细胞上表达增加,HARRT可使其表达水平有所下降.这种表达增加可能与病毒免疫逃逸及CD4＋T细胞的功能抑制有关,HARRT治疗可部分逆转这种免疫紊乱现象.     

HIV感染者外周血CD4+ CD25+ Foxp3+/CD127low/-调节性T细胞表达水平对疾病进展的影响

目的:探讨HIV感染者外周血中CD4+ CD25+Foxp3+调节性T细胞(Treg)、CD4+ CD25+ CD127low/-Treg的水平及其与其他免疫指标的关系.方法:采集68例未经抗HIV治疗的HIV/AIDS患者(长期不进展组即LTNP组29例、典型进展的HIV感染组27例、AIDS组12例)及20例健康成人的外周抗凝全血,经免疫荧光染色,应用流式细胞仪分析CD4+T细胞、CD8+T细胞、NK细胞及CD4+CD25+Foxp3+/CD127low/-Treg的含量,并进行统计学分析.结果:除CD8+T细胞外,HIV/AIDS患者外周血中CD4+T细胞、NK细胞、CD4 +/CD8+结果均明显低于健康对照组(P＜0.05);随着疾病的进展,LTNP组、HIV组、AIDS组CD4+T细胞百分比、绝对值计数,CD8+T细胞绝对计数,NK细胞绝对计数,CD4 +/CD8+比值逐渐下降,而CD8+T细胞百分比逐渐上升.对CD4+ CD25+ Foxp3+ Treg与CD4+ CD25+CD127low/-Treg百分含量、绝对计数进行多重比较发现,各组间CD4+ CD25+ Foxp3+ Treg与CD4-CD25+ CD127low/-Treg所占CD4+T细胞百分含量的差异均有统计学意义(P＜0.05),并且随着疾病的发展,CD4+ CD25+ Foxp3+/CD127low/ Treg细胞百分含量逐渐上升,LTNP组与健康对照组之间、HIV组和AIDS组之间CD4+ CD25+Foxp3+/CD127low/ Treg绝对计数差异无统计学意义(P＞0.05),其余各组间的差异均有统计学意义(P＜0.05),并且随着疾病的发展,CD4+ CD25+ Foxp3-/CD127low/-Treg绝对计数逐渐下降.结论:CD4+ CD25+ Foxp3 +/CD127low/-Treg在HIV持续感染的免疫发病机制中有一定作用.     

尖锐湿疣患者外周血T淋巴细胞上活化抗原的表达

目的 :探讨尖锐湿疣 (CA)患者外周血CD6 9和HLA DR分子在T淋巴细胞上表达的变化及其意义。方法 :采用免疫荧光三标记流式细胞术检测 30例CA患者外周血T细胞CD6 9和HLA DR抗原的表达 ,并以 31例正常人作为对照。结果 :CA患者外周血CD3 T细胞CD6 9的表达 (6 6 3%± 3 13% )与正常人对照组 (5 12 %± 1 6 4 % )相比 ,差异有显著性 (P <0 0 5 ) ,CD4 T细胞CD6 9的表达与正常人对照组相比 ,差异无显著性 (P >0 0 5 ) ,CD8 T细胞表达CD6 9水平 (4 6 1%± 3 0 9% )明显高于对照组 (2 6 7%± 1 31% ,P <0 0 1) ;患者组CD3 T细胞中HLA DR 细胞 (2 1 6 5 %± 8 84 % )比对照组 (13 5 6 %± 5 15 % )显著增高 (P <0 0 0 1)。结论 :CA患者外周血T淋巴细胞的激活以CD8 T细胞为主 ,其免疫激活状态在抗病毒感染中起着重要作用。     

流式细胞术检测CD4~+ T细胞内p24抗原辅助诊断HIV-1的感染

目的探讨流式细胞术(FCM)检测CD4+T淋巴细胞内p24抗原对人免疫缺陷病毒1(HIV-1)感染的辅助诊断价值。方法通过FCM检测HIV-1感染者和正常人(阴性对照)CD4+T淋巴细胞内HIV-1 p24抗原,建立方法。收集HIV-1早期感染者样本,用FCM检测CD4+T淋巴细胞内p24抗原,ELISA检测血浆p24抗原以及nest-PCR检测核酸,比较3种方法的检测结果。结果感染者p24+CD4+T淋巴细胞比例明显高于相应阴性对照(P0.01);感染者组p24+CD4+T淋巴细胞比例95%百分位数为1.92%,确立阴阳界值为2.00%。CD4+T淋巴细胞计数≤350个/μL的感染者p24+CD4+T淋巴细胞比例明显高于相应CD4+T淋巴细胞计数350个/μL的感染者(P0.05)。FCM检测HIV-1早期感染者CD4+T淋巴细胞内p24抗原结果显示,该方法的检验效能优于ELISA检测血浆p24抗原,和核酸检测相当。结论 FCM检测CD4+T淋巴细胞内p24抗原能及时发现HIV-1早期感染,在HIV-1感染的辅助诊断方面有一定的应用价值。     

宫颈癌患者外周血中CD4+CD25+Foxp3+调节性T淋巴细胞及血清中IL-10、TGF-β的表达及其临床意义

目的 探讨宫颈癌患者外周血CD4+ CD25+ Foxp3+调节性T淋巴细胞(regulatory T cells,Treg)、T淋巴细胞亚群以及血清IL-10、TGF-β1、TGF-β2细胞因子的表达及其临床意义.方法 选取32例宫颈癌患者和24位健康体检者为研究对象,采用流式细胞术检测受试者外周血中CD4+ CD25+ Foxp3+ Treg占CD4+T淋巴细胞的比例、T淋巴细胞亚群细胞比例;采用ELISA方法检测血清中细胞因子IL-10、TGF-β1、TGF-β2的含量.结果 与健康对照组相比,宫颈癌组外周血中CD4+CD25+ Foxp3+ Treg占CD4+T淋巴细胞的比值明显增高,差异有统计学意义(P＜0.01),T淋巴细胞亚群(CD3+、CD4+、CD8+、CD4 +/CD8+、CD19+)比例差异无统计学意义(P＞0.05),血清IL-10含量明显增高,TGF-β2含量明显降低,差异有统计学意义(P＜0.01),TGF-β1的含量差异无统计学意义(P＞0.05);宫颈癌组手术后、化疗后CD4+ CD25+ Foxp3+ Treg占CD4+T淋巴细胞的比例,血清IL-10、TGF-β1、TGF-β2含量均明显降低,差异有统计学意义(P＜0.01),CD4+ CD25+ Foxp3+Treg与TGF-β1之间存在正相关(r=0.673,P＜0.01).结论宫颈癌患者CD4+ CD25+ Foxp3+ Treg占CD4+T淋巴细胞的比值增高,且与TGF-B1含量呈正相关,可能在宫颈癌的肿瘤发生发展及肿瘤免疫逃逸中起着重要作用.     

CD8 lymphocytosis in primary cytomegalovirus (CMV) infection of allograft recipients: expansion of an uncommon CD8+ CD57- subset and its progressive replacement by CD8+ CD57+ T cells.

Allograft recipients undergoing cytomegalovirus infection present increased proportions of circulating CD8+ lymphocytes. A longitudinal study of 11 kidney and five liver allograft recipients with primary CMV infection but no other etiological factor of graft dysfunction revealed selective imbalances of peripheral blood CD8+ T cell subsets. Initially, CMV viraemia is associated with elevated CD8+bright T cell numbers and T cell activation. Activation markers fall to normal when viral cultures become negative (before the end of the first month). During the second to sixth month, most (12/16) patients keep up high CD8+ T cell counts (1050-2900 CD8+ cells/mm3), comprising an uncommon CD8+ T cell subset, as 45-73% of CD8+bright lymphocytes were CD3+ and TCR alpha beta+, but were not stained by anti-CD28, CD11b, CD16, CD56, and CD57 antibody. Unexpectedly, CD8+CD57+ T cells, a hallmark of CMV infection, do not appear until the second to sixth month of primary CMV infection, and their numbers increase progressively thereafter. They become the predominant CD8+ T cell subset after 6 months of infection and their persistence for several (up to 4) years is strongly correlated (r = 0.87) with expansion of CD8+ cells. By analysis with MoAbs, there was no bias towards the use of particular TCR-V beta gene families at any time of primary CMV infection. Persistence of CD8 lymphocytosis is thus directly related to the rate of expansion of an uncommon CD8+CD57- subset and its progressive replacement by CD8+CD57+ T cells that are chronically elicited by CMV.     

Inhibition of HIV replication by CD8+ T cells correlates with CD4 counts and clinical stage of disease.

We sought to evaluate the relationship of CD8+ T cell-mediated inhibition of autologous HIV replication in vitro to disease stage in HIV+ individuals. Depletion of CD8+ T cells from peripheral blood lymphocytes of 16 HIV+ subjects increased the percentage of virus-producing cultures from 56% to 81%. CD4+ T cells were purified from 52 HIV+ individuals and cultured alone or in the presence of autologous CD8+ T cells. In 13 (25%) subjects HIV replication was only detected in the absence of CD8+ T cells (inhibition positive); in 26 (50%) viral replication occurred both in the absence and presence of CD8+ cells (inhibition negative). In the remaining 13 (25%) subjects, CD8+ T cell-mediated inhibitory activity could not be evaluated because stimulation of their purified CD4+ T cells did not result in p24 production. In some virus culture-negative individuals, the inability to demonstrate HIV replication was due to the presence of low numbers of CD8+ T cells that co-purified with CD4+ T cells. Detection of inhibitory CD8+ T cells was associated with significantly higher CD4 counts and better clinical status compared with inhibition-negative subjects. These results demonstrate that CD8+ T cell-mediated inhibition of HIV replication correlates with disease stage, and thus may play a role in preventing disease progression. CD8+ T cells did not inhibit autologous HIV replication across a semipermeable membrane. Further, the ability of CD8+ T cells to prevent HIV replication did not correlate with lysis of autologous CD4+ T cells. Thus, CD8+ T cells inhibited autologous HIV replication in vitro through a contact-mediated non-lytic mechanism.     

Proliferation and interleukin 5 production by CD8hi CD57+ T cells

CD8hi CD57+ T cells have previously been described as effector memory T cells with minimal expansion capacity and high susceptibility to activation-induced cell death. In contrast, we demonstrate here that CD8hi CD57+ T cells are capable of rapid expansion using multiple techniques including [(3)H]thymidine uptake, flow cytometric bead-based enumeration and standard haemocytometer counting. Previous reports can be explained by marked inhibition of activation-induced expansion and increased 7-amino-actinomycin D uptake by CD8hi CD57+ T cells following treatment with CFSE, a dye previously used to measure their proliferation, combined with specific media requirements for the growth of this cell subset. The ability of CD8hi CD57+ T cells to further differentiate is highlighted by a distinct cytokine profile late after activation that includes the unexpected release of high levels of interleukin 5. These data indicate that CD8hi CD57+ T cells should not be considered as "end-stage" effector T cells incapable of proliferation, but represent a highly differentiated subset capable of rapid division and exhibiting novel functions separate from their previously described cytotoxic and IFN-gamma responses.     

CD8+ CD28- and CD8+ CD57+ T cells and their role in health and disease

Chronic antigenic stimulation leads to gradual accumulation of late-differentiated, antigen-specific, oligoclonal T cells, particularly within the CD8(+) T-cell compartment. They are characterized by critically shortened telomeres, loss of CD28 and/or gain of CD57 expression and are defined as either CD8(+) CD28(-) or CD8(+) CD57(+) T lymphocytes. There is growing evidence that the CD8(+) CD28(-) (CD8(+) CD57(+)) T-cell population plays a significant role in various diseases or conditions, associated with chronic immune activation such as cancer, chronic intracellular infections, chronic alcoholism, some chronic pulmonary diseases, autoimmune diseases, allogeneic transplantation, as well as has a great influence on age-related changes in the immune system status. CD8(+) CD28(-) (CD8(+) CD57(+)) T-cell population is heterogeneous and composed of various functionally competing (cytotoxic and immunosuppressive) subsets thus the overall effect of CD8(+) CD28(-) (CD8(+) CD57(+)) T-cell-mediated immunity depends on the predominance of a particular subset. Many articles claim that CD8(+) CD28(-) (CD8(+) CD57(+)) T cells have lost their proliferative capacity during process of replicative senescence triggered by repeated antigenic stimulation. However recent data indicate that CD8(+) CD28(-) (CD8(+) CD57(+)) T cells can transiently up-regulate telomerase activity and proliferate under certain stimulation conditions. Similarly, conflicting data is provided regarding CD8(+) CD28(-) (CD8(+) CD57(+)) T-cell sensitivity to apoptosis, finally leading to the conclusion that this T-cell population is also heterogeneous in terms of its apoptotic potential. This review provides a comprehensive approach to the CD8(+) CD28(-) (CD8(+) CD57(+)) T-cell population: we describe in detail its origins, molecular and functional characteristics, subsets, role in various diseases or conditions, associated with persistent antigenic stimulation.     

CD57 expression by T cells in the female genital tract of HIV-zx1 infected women

Despite an influx of T cells to the cervix during HIV infection, genital T cells are not associated with control of HIV shedding. CD57 expression by T cells has been associated with enhanced migratory potential and CD57+ T cells have been shown to accumulate in tissues during the late stages of HIV disease. We investigated the impact of HIV-infection and clinical status on the expression of CD57 by T cells from the female genital tract in 13 HIV-infected and 5 uninfected women. We found that cervical and blood-derived T cells expressed similar frequencies of CD57. The frequency of CD57 expression by cervical or blood T cells was not associated with clinical status (CD4 counts). No impairment in IFN-γ production by CD57+ T cells from the genital tract was observed. We conclude that increased T cell senescence does not appear to be a hallmark of genital mucosal HIV-1 infection.     

Circulating CD4+CD8+ T lymphocytes in patients with Kawasaki disease

We serially monitored cell surface antigen expression on mononuclear cells in peripheral blood isolated from patients with Kawasaki disease (KD), and found, for the first time, that a markedly increased number of CD4⁺CD8⁺ T lymphocytes was present in some of the patients (11 of the 24 cases). The cases of five of these 11 patients were complicated with coronary artery lesion (CAL); the 13 patients with normal numbers of CD4⁺CD8⁺ T lymphocytes did not have CAL. The patients' age, sex and grade of systemic inflammation evaluated by peripheral leucocyte count and serum C-reactive protein levels were not correlated to the number of CD4⁺CD8⁺ T lymphocytes. Other cell surface antigen characteristics of the CD4⁺CD8⁺ T lymphocytes included CD3⁺, CD45RA⁺, CD45RO⁺, CD16^?, and HLA-DR⁺. These results indicate that the surface antigen characteristics of the KD peripheral blood examined were the same as those of Epstein–Barr virus infection without CD45RA⁺. These findings provide useful information for the analysis of the pathogenesis of KD.     

CD57+ T lymphocytes are derived from CD57− precursors by differentiation occurring in late immune responses

CD3⁺ T cells expressing the 110-kDa CD57 antigen are found in survivors of renal, cardiac and bone marrow transplants, in patients with acquired immune deficiency syndrome and in patients with rheumatoid arthritis. They are also present in normal individuals and expand upon ageing. They do not grow in culture and their role in the immune response is poorly understood. The expression of the various isoforms of the leukocyte common antigen (CD45) identifies a spectrum of differentiation in CD4⁺ and CD8⁺ T cells ranging from naive (CD45RA⁺CD45RB^brightCD45RO^?) through early primed cells (CD45RA^?RB^brightRO^dull) to highly differentiated memory cells which are CD45RA^?RB^dullRO^bright. CD45 isoforms expressed by CD57⁺ T cells showed distinct differences between CD4⁺ and CD8⁺ populations, but in each case indicated an advanced state of differentiation. The expression of T cell receptor Vβ families was highly variable between individuals, but both CD57⁺ and CD57^? cells show a full range of the specificities tested. Vβ expression was more closely related within either the CD4⁺ or the CD8⁺ subsets, irrespective of CD57 expression, than between these subsets, suggesting a relationship between CD57⁺ and CD57^? cells within the same T cell pool. This possibility was supported by experiments showing that CD3⁺CD57⁺ lymphocytes were similar to CD3⁺CD57^? T cells in terms of the production of basic T cell cytokines [interleukin (IL)-2, IL-4, and interferon-γ]. Furthermore, in vitro stimulation of CD3⁺CD57^? T cells in secondary mixed leukocyte reaction or by co-culture with IL-2 and IL-4 induced the appearance of CD3⁺CD57⁺ cells with phenotypic and functional similarities to in vivo CD3⁺CD57⁺ cells. These data strongly suggest that the expression of CD57 is a differentiation event which occurs on CD57^? T cells late in the immune response.     

Ex vivo analysis of phenotype and TCR usage in relation to CD45 isoform expression on cytomegalovirus-specific CD8+ T lymphocytes

Human cytomegalovirus (CMV) is a ubiquitous pathogen which sets up a lifelong persistent infection and which can lead to significant disease in the immunosuppressed. The immunological mechanisms controlling CMV in the long term are not defined completely, but CD8+ T lymphocytes are thought to play an important role. Antiviral CD8+ T lymphocytes may exist in very large pools in healthy individuals. Although the detailed composition of these pools is not completely understood, there is known to be heterogeneity, in particular of CD45 isoform expression. We have therefore investigated the CD8+ T-lymphocyte response against CMV directly ex vivo using Class I tetramers combined with stains for a range of phenotypic markers followed by four-colour flow cytometric analysis. In particular, we examined expression of these phenotypic markers in relation to the expression of CD45 isoforms. We found that a spectrum of phenotypes exists stably, from CD45R0(high)/RA(low) through CD45RA(high)/R0(low), and that expression of other surface markers such as CD28 and CD62L, and also TCR usage, may vary in parallel with CD45 isoform expression. In some individuals, expansions of antigen-specific CD8+ T lymphocytes bearing specific TCR Vbeta chains were restricted to cells of particular CD45 isoforms. Immunity against CMV comprises a large population of CD8+ T lymphocytes with heterogeneous potential, a spectrum in which CD45 isoform expression may play a central role.     

Characterization of bronchoalveolar lavage T cell subsets in sarcoidosis on the basis of CD57, CD4 and CD8

T cells expressing CD57 (a natural killer cell marker) with interferon-gamma (IFN-gamma) producing capacity increase under various conditions. CD57+ T cells are also present in the bronchoalveolar lavage fluid (BALF) of sarcoidosis, and several phenotypical and functional analyses of these cells have been reported. In the present study, BALF T cells obtained from 52 patients with sarcoidosis were classified further into CD4+CD57+ T cells, CD4+CD57- T cells, CD8+CD57+ T cells and CD8+CD57- T cells and their phenotypes and functional characteristics were assessed. Substantial proportions of these T cell subsets expressed natural killer cell markers CD161 and CD122. The biased expansion of Vbeta2 T cells was observed in both CD4+CD57+ T cells and CD4+CD57- T cells in BALF from most patients, while the expansion of other Vbeta T cells was also observed in some patients. Unexpectedly, the biased expansion of certain Vbeta T cells was also seen in either CD8+CD57+ T cells or CD8+CD57- T cells, while the expanded Vbeta T cells in CD8+ T cells differed substantially among individuals. BALF T cells showed a remarkably lower T cell receptor (TCR) intensity than that of peripheral blood T cells. Both CD8+ T cell subsets in BALF of sarcoidosis expressed the intracellular perforin/granzyme B, while all four subsets expressed intracellular IFN-gamma after in vitro activation, and CD4+ T cells, especially CD4+CD57+ T cells, expressed tumour necrosis factor-alpha. These findings indicate that CD57+ T cells as well as CD57- T cells in the BALF are phenotypically and functionally different from peripheral blood T cells and may play an important role in the Th1 dominant state and inflammation in pulmonary sarcoidosis.