HIV/AIDS与肝炎病毒感染者CD4+、CD8+淋巴细胞数的比较

目的：探讨ＨＩＶ携带者或ＡＩＤＳ病人（简称ＨＩＶ／ＡＩＤＳ）与肝炎病毒感染者的ＣＤ４＾＋、ＣＤ８＾＋淋巴细胞变化,并进行比较。方法：用免疫磁珠法（ＤＹＮＡ ｂｅａｄｓ）检测ＣＤ４＾＋、ＣＤ８＾＋Ｔ淋巴细胞数,用双侧ｔ检验进行统计学处理。结果：全部ＨＩＶ／ＡＩＤＳ的ＣＤ４＾＋Ｔ淋巴细胞降低,且幅度大,８３．４％低于５００细胞／μｌ。在病毒性肝炎中降低的百分比和幅度均明显低于前者,只有２８．８％低于５     

存放温度及时间对于HIV/AIDS患者外周血CD4+、CD8+细胞测定结果的影响

目的　研究HIV AIDS患者外周血CD4 、CD8 淋巴细胞数在不同条件下 (时间、温度和处理过程 )的变化。方法　选取HIV AIDS患者 34例 ,用流式细胞术检测在 4℃条件下放置不同时间(2、2 4、4 8、72h)的外周血CD4 、CD8 细胞数的变化 ,对经过处理的外周血 (处理过的血样 )CD4 、CD8 细胞数的变化进行比较 ;对室温条件下放置不同时间 (2、2 4、4 8、72h)处理过的血样CD4 、CD8 细胞数的变化进行比较。结果　在 4℃时 ,全血放置 2、2 4、4 8、72h的CD4 细胞计数差异无显著意义(P >0 0 5 ) ,而CD8 细胞数放置 72h时则差异有显著意义 (P <0 0 5 ) ;处理过的样品放置 72hCD4 细胞计数差异才有显著意义 (P <0 0 5 ) ,而CD8 细胞数在 2 4h时差异就有显著意义 (P <0 0 5 )。在室温时 ,处理过血样放置 2、2 4、4 8、72h的CD4 细胞计数差异无显著意义 (P >0 0 5 ) ,而CD8 细胞数在 4 8h则差异有显著意义 (P <0 0 5 )。结论　抗凝全血在 4℃放置 4 8h ,检测CD4 、CD8 淋巴细胞数 ,结果是可靠的。处理过血样在室温放置 2 4h ,检测CD4 、CD8 淋巴细胞数 ,结果是可靠的。 2 4～4 8h虽然CD4 淋巴细胞没有变化 ,但CD8 淋巴细胞却发生明显的变化 ,两者比例必然发生变化。     

中国HIV/AIDS患者NK细胞及NKT细胞变化的检测

目的　探讨HIV感染后机体NK(naturalkillercells)及NKT细胞的变化情况。方法　取外周血细胞 ,用标记荧光的抗体进行染色 ,流式细胞仪分析HIV AIDS患者NK和NKT细胞的变化。结果　HIV AIDS患者NK、NKT细胞和CD4 + T细胞显著低于正常对照 ;CD8+ T细胞显著高于正常对照。HIV AIDS患者NK百分数显著低于正常对照 ,与CD4 + T细胞数量成正比 ,r=0 .2 89,P <0 .0 1 ;NKT细胞数量与CD4 + T细胞数量成正比 ,r =0 .378,P <0 .0 1 ;与CD8+ T细胞数量成正比 ,r =0 .340 ,P <0 .0 1 ;长期不进展组NKT、NK细胞数量与正常对照组差异无显著性。结论　HIV感染可明显降低HIV AIDS患者NK和NKT细胞数量 ,NK和NKT细胞变化与疾病进展密切相关。     

我国HIV/AIDS患者外周血CD4+T淋巴细胞凋亡与疾病进展的相关性分析

HIV特征性地引起CD4^＋T细胞逐步减少和进行性免疫缺陷。尽管目前HIV导致CD4^＋T细胞死亡的具体机制仍不完全清楚，但越来越多证据支持HIV引发的淋巴细胞凋亡是一个重要因素。有研究显示HIV感染使CD4^＋T细胞上多种凋亡相关分子表达异常，但有关CD4^＋T细胞凋亡率与疾病进程的报道较少，更少见到中国HIv／AIDS患者CD4^＋T细胞凋亡在不同疾病进程中变化特点的研究资料。本实验采用流式细胞术研究了我国HIV／AIDS患者CD4^＋T细胞凋亡情况及其在不同疾病阶段中的变化特点。     

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...     

HIV/AIDS患者特异性细胞毒性T细胞功能的研究

目的 了解中国HIV／AIDS患者HIV特异性细胞毒性T细胞(CTL)功能。方法 将覆盖HIV-1 P15、P17和P24 Gag全长的94个重叠多肽作为抗原，用IFN-γ ELISPOT方法检测HIV／AIDS患者HIV-1特异性CTL功能。结果 HIV-1抗原多肽P17-15、P17-16、P24-7、P17-8，P24-28最易被HIV／AIDS患者特异性CTL识别。HIV感染者识别HIV-1多肽的数量和强度均高于AIDS患者。结论 我国HTV／AIDS患者体内存在识别不同HIV-1 Gag多肽的特异性CTL，且HIV特异性CTL功能与疾病进展相关。     

广东HIV/AIDS患者舌象分析及其与CD4、CD8、CD4/CD8计数的相关性研究

目的分析广东HIV/AIDS患者的舌象规律及其与CD4、CD8、CD4/CD8计数的规律。方法对609例广东HIV/AIDS患者进行舌象分析和CD4、CD8T淋巴细胞检测,探讨其相关性。结果HIV/AIDS患者CD4、CD4/CD8比值均低于正常水平;两组淡红舌患者的CD4、CD4/CD8比值均高于其他4种舌色(淡白舌、红舌、暗舌、紫舌)(P<0.05);两组淡白舌患者的CD4、CD4/CD8比值均低于其他4种舌色(P<0.05);HIV/AIDS患者暗舌的比例(41.38%)均大于其他舌色,其次是红舌(35.47%)(P<0.05);HIV/AIDS患者以白腻苔(44.83%)为主,其次为黄腻苔(26.11%)、薄白苔(20.20%),少苔(3.45%)。结论舌象能客观反映HIV/AIDS患者身体免疫机能;广东HIV/AIDS患者多出现正气虚,夹瘀、夹痰化热的表现。     

RA患者外周血CD8＋CD28-和CD4＋CD25＋调节性T细胞的表达及疾病活动性指标相关性分析

目的：检测类风湿性关节炎（RA）患者外周血CD8＋CD28-、CD4＋CD25＋调节性T细胞亚群,探讨其与临床活动性指标的关系。方法：采用流式细胞术检测台州医院RA患者外周血CD8＋CD28-、CD4＋CD25＋ T细胞亚群比例,探讨调节性T细胞与RA活动性、类风湿因子（RF）、免疫球蛋白（Ig）、C反应蛋白（CRP）、补体C3、抗CCP抗体、抗核抗体（ANA）、血小板（PLT）及血沉（ESR）的关系。结果：活动期RA患者外周血CD4＋CD25＋调节性T细胞亚群比例显著低于正常对照组（P〈0.01）,但稳定期RA患者与正常对照组结果差异无统计学意义（P〉0.05）。活动期和稳定期RA患者CD8＋CD28-与正常对照组相比较,结果无统计学意义（P〉0.05）;CD4＋CD25＋与CRP密切相关（r=-0.593,P〈0.05）,CD8＋CD28-与ESR相关系数呈弱相关。CD4＋CD25＋和CD8＋CD28-细胞与RF、IGG、C3、ANA、anti-CCP和PLT未见明显相关性。结论：活动期RA患者外周血CD4＋CD25＋ T细胞亚群比例减少,CD4＋CD25＋ T细胞可能与类风湿性关节炎疾病进展有关。     

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。     

膈下迷走神经切断对外周血CD4+/CD8+T细胞比值的影响

近年来越来越多的证据显示 ,神经系统和免疫系统存在着密切的相互影响。免疫系统在接受抗原刺激迅速作出反应的同时 ,还将免疫信息传递到中枢神经系统 ,影响该系统的活动并接受其调节。目前 ,对免疫信息向中枢传递的途径有几种假说 ,其中受到重视的是神经机制 ,特别是迷走神经可能起重要的作用。免疫信息可能通过刺激迷走神经感觉神经纤维或旁神经节细胞引起电活动将免疫信息上传至脑。本研究中我们切断大鼠膈下迷走神经后 ,通过观察外周血ＣＤ4 /ＣＤ8 Ｔ细胞比值的改变 ,探讨了迷走神经在神经系统调节免疫系统功能的环路中所起的作用。1…     

A subset of functional effector-memory CD8+ T lymphocytes in human immunodeficiency virus-infected patients

CD8(+) T cells provide protective immune responses via both cytolytic and non-cytolytic mechanisms in subjects infected with human immunodeficiency virus (HIV). In the present study, we investigated the CD28 expression of CD8(+) T cells present in the peripheral blood lymphocyte subset isolated from chronically HIV-infected subjects. Using flow cytometric analysis, a continuous spectrum of CD28 intensity ranging from negative to high, which could be separated into CD28-negative, intermediate (int) and high, was seen for CD8(+) T cells. Our study focused mostly on the CD28(int) CD8(+) T cells. The CD28(int) CD8(+) T cells are CD57(-) CD27(+) CD45RO(+) CD45RA(-) CCR7(low) CD62L(int). The proliferative capacity of CD28(int) CD8(+) T cells was intermediate between those of CD28(-) CD8(+) T cells and CD28(high) CD8(+) T cells. The CD28(int) CD8(+) T cells are specific for HIV, cytomegalovirus (CMV) and Epstein-Barr virus (EBV) antigens as measured by human leucocyte antigen pentamer binding and produce both intracellular interferon-gamma and tumour necrosis factor-alpha in response to their cognate viral peptides. The CD28(int) CD8(+) T cells have HIV-specific, CMV-specific and EBV-specific cytotoxic activity in response to their cognate viral peptides. These findings indicate that a subset of functional effector-memory CD8(+) T cells specific for HIV, CMV and EBV antigens may contribute to an efficient immune response in HIV-infected subjects.     

The CD28/HLA-DR expressions on CD4+T but not CD8+T cells are significant predictors for progression to AIDS

To investigate the changes of CD28 and HLA-DR molecules on CD4+ and CD8+ T cells during HIV infection, we classified 130 HIV-infected Koreans into four groups by the CD4 level as follows: group I (> or = 500 cells/mm3), group II (201-499 cells/mm3), group III (51-200 cells/mm3), and group IV (< or = 50 cells/mm3). In CD4+ T cells, the proportion of CD28 expression decreased significantly with the CD4 level while the proportion of HLA-DR expression increased gradually. In particular, the changes of HLA-DR expressions on CD4+ T cells were parallel to the loss of CD28 molecules from stage III to IV. However, the CD28 expression on CD8+ T cells decreased dramatically in the early stage of HIV infection, and the sum and pattern of CD28 and HLA-DR expressions on CD8+ T cells was stable after the first stage. Even though CD28 down-regulation on CD8+ T cells was very severe from the early stage of HIV infection, it might not influence the survival time of HIV-infected Koreans. The sum of the CD28+ subsets and HLA-DR subsets in each T cell was stable in all stages of disease progression. The sums of the CD28+ subsets and HLA-DR+ subsets in CD4+ T and CD8+ T cells were constant as approximately 100% and 55-60% of each T cell. These results suggested that the changes of CD28/HLA-DR expressions on CD4+ T cells were more predictable than those on CD8+ T cells in the evaluation of the disease progression during HIV-infected periods. However, we need further studies to understand why the sum of two molecules in each T cell are constant.     

Ligation of either CD2 or CD28 rescues CD4+ T cells from HIV-gp120-induced apoptosis

Temporal or quantitative imbalance in signals delivered to T cells via T cell antigen receptor (TCR), the CD4 co-receptor, and accessory molecules can lead to anergy, apoptosis, or both. This has been observed following ligation of CD4 by HIV gp120 prior to TCR occupancy. The ability of molecules such as CD2 and CD28, interacting with their ligands LFA-3 and B7, to provide signals that protect T cells from the induction of anergy, has been reported. Here, we demonstrate that ligation of CD2 and CD28 in conjunction with TCR occupancy rescue T cells that have been programmed for apoptotic death by prior CD4 ligation to gp120. This appears to be the result of augmented interleukin-2 and interleukin-4 release by the T cells following these molecular interactions. In conclusion, our results suggest that an impairment of antigen-presenting accessory cell functions could favor gp120-mediated apoptosis in HIV-uninfected cells.     

用流式细胞仪检测HIV感染者和AIDS患者的T细胞亚群

目的用流式细胞仪（ｆｌｏｗｃｙｔｏｍｅｔｅｒ,ＦＣＭ）检测周围血中ＣＤ４＋、ＣＤ８＋淋巴细胞,结合临床情况对ＨＩＶ感染者和ＡＩＤＳ患者的免疫状况进行评价。方法将抗凝全血进行白细胞分类计数,用双色荧光标记的单克隆抗体染色,经溶血和固定后,用ＦＣＭ计数,从而得出ＣＤ４＋、ＣＤ８＋淋巴细胞数。结果ＨＩＶ感染者和ＡＩＤＳ患者的ＣＤ４＋淋巴细胞数都比正常人低,特别是ＡＩＤＳ患者,他们的ＣＤ４＋淋巴细胞数都低于２００个／ｍｍ３,临床表现也很差。结论ＦＣＭ检测结果与临床评价高度一致,而且ＦＣＭ比一般人工计数法更准确、方便、迅速,同时也证明ＦＣＭ是监测ＨＩＶ感染者和ＡＩＤＳ患者的免疫状况的最佳方法。     

Deletion of T lymphocytes in human CD4 transgenic mice induced by HIV-gp120 and gp120-specific antibodies from AIDS patients

CD4, a T cell receptor for major histocompatibility complex class II antigen, is a key regulator of immunological reactivities. When engaged together with the T cell antigen receptor, CD4 enhances immune reactions, whereas when ligated independently of the antigen receptor CD4 inhibits the activation of T cells or initiates their deletion. CD4 serves also as a receptor for the human immunodeficiency virus (HIV), which binds the receptor with high avidity through its envelope molecule, gp120. Studies in tissue culture have shown that its affinity to CD4 gives the virus opportunities to utilize CD4-mediated signaling and to manipulate immunocytes. We show here in human CD4 transgenic mice that appropriately cross-linked HIV envelope protein causes massive deletion of HIV-reactive T cells in vivo.     

慢性乙型肝炎患者外周血CD4＋CD25＋、CD8＋CD28＋淋巴细胞的变化及与病毒载量的关系

目的：研究慢性乙型肝炎患者外周血CD4＋CD25＋、CD8＋CD28＋淋巴细胞的表达变化及与病毒载量的关系。方法：利用流式细胞术检测50例慢性乙型肝炎患者外周血CD4＋CD25＋、CD8＋CD28＋淋巴细胞的表达率和绝对数,并与正常对照组30例进行比较;荧光定量PCR检测慢性乙型肝炎患者HBV核酸的载量,并与CD4＋CD25＋、CD8＋CD28＋淋巴细胞的表达进行相关分析。结果：慢性乙型肝炎患者的CD4＋CD25＋表达率和细胞数分别为（15.60±5.86）%和（0.34±0.13）×109/L,明显高于正常对照组（P〈0.01）;而CD4＋CD25-、CD8＋CD28＋为（21.13±5.32）%、（0.47±0.19）×109/L和（9.49±2.57）%、（0.21±0.07）×109/L,均低于正常对照组（均P〈0.01）;不同病毒载量的慢性乙型肝炎患者CD4＋CD25＋T细胞与病毒载量均呈正相关（r分别为0.552、0.588,P均〈0.01）,而CD8＋CD28＋T细胞与病毒载量均无相关性（r分别0.275、-0.092,P均〉0.05）。结论：慢性乙型肝炎患者细胞毒T细胞（CD8＋CD28＋）减少,免疫调节细胞的增加与慢性乙型肝炎患者病毒长期存在,病程迁延不愈有关。     

SLE患者Th1/Th2平衡状态及与外周血淋巴细胞CD28及CTLA-4分子表达的关系

目的:了解SLE患者Th1/Th2平衡状态以及共刺激分子CD28/CTLA-4与Th1/Th2平衡状态的关系。方法:研究对象为18例SLE患者(活跃期12例、缓解期6例)。对照组14例,为健康体检者。外周血单个核细胞(PBMCs)经梯度密度离心法分离后置于含PMA(5μg/L)及ionomycin(500μg/L)培养液中培养72 h。采用ELISA方法检测培养的PBMCs上清液中IFN-γ及IL-10的含量。应用流式细胞技术检测培养的淋巴细胞CD28及CTLA-4分子的表达。结果:活跃期SLE患者培养的PBMCs分泌IL-10的量(351.29 ng/L±153.31 ng/L)较对照组(254.48 ng/L±120.69 ng/L)有一定程度的升高,但差异无显著(P0.05),IFN-γ的分泌量(25.76 ng/L±16.09 ng/L)明显低于对照组(50.71 ng/L±27.92 ng/L,P0.05),IL-10/IFN-γ比值(18.74±13.77)明显高于对照组(6.66±4.95,P0.05)。培养前、后SLE患者CD3+及CD8+T细胞CD28分子表达量与对照组比较均无显著差异。培养前活跃期SLE患者CD3+T细胞CTLA-4分子表达量(0.79%+0.37%)较对照组(1.31%+0.61%)明显降低(P0.05)。培养后SLE患者CD3+T细胞及CD8+T细胞CTLA-4分子表达量仍低于对照组,但差异无显著(P0.05)。活跃期SLE患者培养的PBMCs中CD3+T细胞CTLA-4分子的表达量与上清液中IFN-γ含量呈明显的直线正相关关系(r=0.681,P0.05)、与上清液中IL-10及IL-10/IFN-γ比值呈明显的直线负相关关系(r=-0.624,P0.05;r=-0.738,P0.01)。结论:SLE患者存在Th1/Th2平衡向Th2方向偏移,即Th2优势状态。CTLA-4分子可能通过抑制CD28的信号转导参与Th2优势状态的形成。     

Evidence for intact costimulation via CD28 and CD27 molecules in hyporesponsive T cells from human immunodeficiency virus-infected individuals

In the activation of T cells, the primary signal is antigen-specific and given through T cell receptor (TcR)/CD3 ligation. Furthermore, costimulatory molecules such as CD28 and CD27, provide an essential signal for activation through interaction with their ligands, present on the membrane of antigen-presenting cells. During asymptomatic human immunodeficiency virus (HIV)-1 infection, T cell function is progressively lost. Here, we investigated whether in the presence of impaired responses of T cells from HIV-infected individuals to signal one, costimulation through CD28 and CD27 after interaction with their natural ligands CD80 and CD70 is intact. T cell proliferative responses to signal one in combination with CD80 or CD70 were decreased in a large fraction of asymptomatically HIV-infected individuals. This was due to impaired responses of signal one but not to impaired responses to costimulation, since CD80 or CD70 did enhance signal one-mediated proliferative responses to a normal extent. Moreover, in individuals with proliferative responses to signal one that were decreased to 50% of normal T cell responses, costimulation even was increased compared to controls. Our results demonstrate that in HIV-infected individuals the response to costimulation is relatively preserved compared to responses to the first signal and point to the defect in T cells in HIV infection being primarily in the CD3/TcR-mediated pathway.     

CD8+T淋巴细胞非细胞毒性抗HIV作用研究进展

细胞免疫在抗病毒感染中发挥着至关重要的作用。人类免疫缺陷病毒(HIV)感染所引起的细胞免疫反应是由CD8 T淋巴细胞的亚群细胞毒性T细胞(CTL)介导的先天免疫。CTLs一方面通过细胞毒性作用杀伤感染的细胞,另一方面分泌可溶性抗病毒因子(CAF)发挥直接的抗病毒作用,因此成为HIV感染中细胞免疫的重要组成部分。本文就HIV感染这一疾病过程中CD8 T淋巴细胞所发挥的非细胞毒性抗病毒效应作一概述。