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     

献血员CD8^＋T细胞的增另对CD4^＋T细胞的抑制作用

实验发现６５％体检合格的献血员外周血ＣＤ８＾＋Ｔ细胞数量明显增加,并伴随ＣＤ１６＾＋淋巴细胞数量的增加。这些异常的淋巴细胞与葡萄球菌肠毒素Ｂ（ＳＥＢ）共同２６ｄ,ＣＤ４＾＋Ｔ细胞无增殖反应。预先用抗ＣＤ８抗体去除ＣＤ８＾＋Ｔ细胞（去除率〉９０％）再用ＳＥＢ刺激淋巴细胞,其应答能力恢复正常,增殖的细胞是ＣＤ４＾＋Ｔ细胞,它们由３４．０４％增加到９９．３４％。ＣＤ８＾＋Ｔ细胞由最初的９．５７％降低到０     

CD8^＋的树突状细胞研究进展

ＣＤ８＾＋的树突状细胞是树突状细胞中的一个特殊的群体,可能主要来源于胸腺前体细胞,其主要特点是高度表达ＣＤ８分子和一些Ｔ细胞的表面标记,在功能上主要表现为对ＣＤ４＾＋Ｔ细胞的活化的同时,又介导其凋亡;而对ＣＤ８＾＋Ｔ细胞的作用是通过抑制内源性ＩＬ－２的产生从而抑制其生物学作用。     

非细胞毒性CD8^＋T细胞应答—控制HIV感染的有效途径

以往一直认为，ＨＩＶ感染者体内的细胞免疫主要是由细胞毒性ＣＤ８＾＋Ｔ细胞介导，导致病毒感染细胞ＣＤ４＾＋细胞的裂解或凋亡。但近年来发现，非细胞毒性ＣＤ８＾＋Ｔ细腻可在不杀伤感染细胞的情况下有效控制ＨＩＶ的感染。     

雷公藤抑制致敏小鼠CD4^＋T细胞核因子GATA3,NFAT的活性

目的 探讨致敏小鼠ＣＤ４＾＋Ｔ淋巴细胞核转录因子ＧＡＴＡ３、ＮＦＡＴ活性的变化及雷公藤内酯醇（ＴＰ）的作用。方法 采用卵蛋白（ＯＶＡ）致敏的方法建立模型；运用ｐａｎｎｉｎｇ法分离ＣＤ４＾＋Ｔ淋巴细胞；通过凝胶电泳迁移率实验（ＥＭＳＡ）对ＣＤ４＾＋Ｔ淋巴细胞核因子ＧＡＴＡ３、ＮＦＡＴ的ＤＮＡ结合活性及ＴＰ的作用进行检测，同时就ＴＰ的作用与地塞米松（ＤＭ）相比较。结果 正常小鼠ＣＤ４＾＋Ｔ淋巴细胞核因     

超抗原SEB激活诱导CD4^＋T细胞凋亡模型的建立

目的 为了探讨超抗原活化诱导ＣＤ４＾＋Ｔ淋巴细胞凋亡分子机理及其信号传导途径,有必要建立超抗原活化诱导ＣＤ４＾＋Ｔ细胞凋亡模型。方法 采用电镜观察细胞凋亡的形态学特征,借助流式细胞仪ＰＩ染色观察细胞凋亡的光散射特征及亚二倍体核型峰特征,琼脂糖凝胶电泳分析细胞凋亡的ＤＮＡ片段化图谱,最后采用改进的二苯胺（ＤＰＡ）法定量分析细胞凋亡ＤＮＡ片段化百分率。结果 ４ｕｇ／ｍＬ ＳＥＢ刺激静息的ＳＥＢ应答ＣＤ４＾＋Ｔ细胞１２ｈ后,电镜下观察呈现典型的凋亡形态学特征;流式细胞仪ＰＩ染色分析表明,在二倍体峰的左侧出现亚二倍体核型峰典型凋亡特征,在光散射图谱上呈现低于正常细胞的前向散射和高于正常细胞的侧向散射;琼脂糖凝胶电泳分析呈现典型的凋亡ＤＮＡ梯状图谱;ＤＮＡ片段化百分率分析表明,超抗原ＳＥＢ活化诱导ＣＤ４＾＋Ｔ细胞凋亡率的     

分泌细胞因子的CD8^＋T细胞亚群研究进展

Ｔ细胞具有功能不同的亚群，其中ＣＤ４＾＋辅助Ｔ细胞（ＴＨ）的亚群分类及功能早已成定论。近几年的研究发现传统的ＣＤ８＾＋细胞毒Ｔ细胞（ＣＴＬ）也可分泌与ＣＤ４＾＋Ｔ细胞相似的细胞因子，也可分为两类亚群，分别定义为Ｔｃ１和Ｔｃ２。本文就近年来这方面的研究进展作一综述。     

CD8T细胞转换成能产生TH2细胞因子并辅助B细胞的非溶细胞CD8—CD4—…

ＣＤ８＋Ｔ细胞是抵抗病毒和胞内寄生茵感染的主要防御机构。其干扰－ｒ（ＩＦＮ－ｒ）产生能力以及其溶细胞活性是对这些病原体的免疫应答的关键性因素。在ＩＬ－４存在的情况下，小鼠的成熟ＣＤ８＋Ｔ细胞性并且不产ＩＦＮ－ｒ的ＣＤ８＾－ＣＤ４＾－细胞群。可是，这些ＣＤ８＾－细胞能产生大量的ＩＬ－４，ＩＬ－５和ＩＬ－１０，并能辅助激活静止的Ｂ细胞。因而，ＣＤ８效应机能机能是多样化的。     

CD4^＋T细胞的抗瘤作用

ＣＤ４＾＋Ｔ细胞在机体抗瘤免疫反应中的作用来越来越受到重视，ＣＤ４＾＋Ｔ细胞不仅可以分泌大量的细胞因子和辅助ＣＤ８＾＋Ｔ细胞杀伤肿瘤细胞，而且ＣＤ４＾＋Ｔ细胞在肿瘤免疫中具有免疫记忆和直接杀伤肿瘤细胞的功能，因而机体的抗瘤免疫反应中起重要作用。过继转输ＣＤ４＾＋Ｔ细胞是肿瘤过继免疫治疗的一个新方案。     

不同胃病患者外周血T淋巴细胞亚群的测定结果

应用流式细胞仪（ＦＣＭ）检测外周血Ｔ淋巴细胞亚群ＣＤ３＾＋，ＣＤ４＾＋、ＣＤ８＾＋及ＣＤ４＾＋／ＣＤ８＾＋比值。健康人（志愿献血者）２０例，消化性溃疡３０例，萎缩性胃炎３０例，慢性浅表性胃炎３０例。结果表明：消化性溃疡、萎缩性胃炎患者Ｔ淋巴细胞亚群ＣＤ３＾＋、ＣＤ４＾＋均值明显低于健康人（Ｐ〈０．０１）。消化性溃疡患者ＣＤ＾８＋明显高于健康人（Ｐ〈０．０５），浅表性胃炎患者与健康人比Ｔ细胞亚群无明     

Preferential S phase entry and apoptosis of CD4(+) T lymphocytes of HIV-1-infected patients after in vitro cultivation

We have studied the relationship between spontaneous apoptosis and cell cycle perturbations in circulating peripheral blood lymphocytes of HIV-1-infected patients and healthy controls. PBMC obtained from HIV-1-infected patients and healthy controls were incubated in culture medium for 48 h. Cells were separated into CD4(+) and CD8(+) populations using immunomagnetic beads. Apoptosis and cell cycle phases were measured by propidium iodide staining and bromodeoxyuridine (BrdU) incorporation followed by flow cytometric analyses. In experiments using cells obtained from HIV-1-infected patients, spontaneous apoptosis was more frequent in CD4(+) T lymphocytes than in CD8(+) T lymphocytes (17.6% vs 9.5%, P < 0.005). Among healthy controls, spontaneous apoptosis in CD4(+) and CD8(+) T lymphocytes was comparable (4.5% vs 5.1%). Lymphocytes obtained from patients were more frequently in S phase than healthy controls' cells (2.2 +/- 0.9% vs 0.5 +/- 0.2%, P < 0.002) and patients' CD4(+) cells tended to enter S phase more frequently than controls' CD4(+) cells (4.2% +/- 3.5% vs 1.8% +/- 0.5% P < 0.04), whereas the frequency of S phase CD8(+) T cells was not different among patients (2.8% +/- 2.9%) and controls (1.8% +/- 0.5%) (P > 0.4). Kinetic analyses using BrdU and PI staining revealed that S phase cells were more likely to become apoptotic than resting (G(0)-G(1)) cells (28.4% +/- 10.3% vs 11.3% +/- 9.9% in patients, P < 0.04, and 15.3% +/- 2.8% vs 1.8% +/- 0.5% in controls, P < 0.003). Lymphocytes obtained from HIV-1-infected persons are activated in vivo to enter S phase and to undergo spontaneous apoptosis after brief in vitro cultivation. The present studies indicate that most apoptotic cells in this system are CD4(+) and kinetic analyses reveal that S phase cells are more likely to undergo spontaneous apoptosis than G(0)-G(1) cells. Accelerated cell death in HIV-1 disease may contribute to the failure of lymphocyte responsiveness to appropriate T cell receptor stimulation.     

类风湿关节炎CD4+CD28-T细胞与淋巴细胞凋亡的相关性研究

目的: 研究类风湿关节炎(RA)患者外周血CD4⁺CD28^-T细胞比例与淋巴细胞凋亡异常的相关性。方法: 采用流式细胞术三色分析法检测50例患者和50例健康志愿者的外周血淋巴细胞中CD4⁺CD28^-T细胞比例;通过加入PHA孵育检测RA病人外周淋巴细胞和正常对照的淋巴细胞对激活诱导细胞死亡(AICD)易感性差异;分析CD4⁺CD28^-T细胞比例与外周血淋巴细胞凋亡率的相关性。结果: RA组CD4⁺CD28^-T细胞比例的均数明显高于健康对照组(7.79%±3.52% vs 1.89%±1.78%,P<0.05)。RA组病人外周血淋巴细胞的AICD凋亡率低于健康对照组(11.38%±5.73% vs 19.46%±6.32%,P<0.05)。Spearman相关分析结果显示CD4⁺CD28^-T细胞比例与外周血淋巴细胞AICD凋亡率负相关(r=-0.433,P<0.01)。结论: RA患者外周血中CD4⁺CD28^-T细胞比例增多,活化淋巴细胞生存期延长,这可能参与RA的发病机制。     

Age-dependent changes in the susceptibility to apoptosis of peripheral blood CD4+ and CD8+ T lymphocytes with virgin or memory phenotype

Susceptibility to apoptosis changes with age and most of the available data on lymphocytes refer to mitogen stimulated cells. We studied this susceptibility in quiescent, purified CD4+ or CD8+ T cells from a group of Italian old people compared with a group of young people. We found that an apoptotic agent such as 2-deoxy-D-ribose (dRib), which acts via glutathione depletion and oxidative stress, was more effective in CD4+ T cells from young donors, while no difference was found in CD8+ T cells. On the contrary, another agent such as TNF-alpha, which acts via receptor engagement, was more effective in CD8+ T cells from old subjects, and no difference was found in CD4+ T cells. When marker of activation-memory were investigated, no difference between young and old subjects was found when dRib was used. Differently, when TNF-alpha was used, memory and activated CD4+ T cells from old donors were less sensitive than younger counterparts, while memory CD8+ T cells from old donors were more sensitive than younger counterparts. This suggests that age-related changes in susceptibility to apoptosis of resting T cells largely depend on the type of the apoptotic stimulus which is used as well as on the memory phenotype of the cells. These results may also account, at least in part, for the deep remodelling of T cell repertoire that occurs during ageing.     

Participation of RANTES and T-cell apoptosis in human renal allograft

The aim of this study was to evaluate the serum RANTES (Regulated upon Activation of Normal T cell Expressed and Secreted) levels and the expression of CCR5, as well as the percentage of apoptotic cells, in peripheral T lymphocytes from renal transplanted patients with acute rejection (AR), chronic rejection (CR) or stable evolution (SE). RANTES serum levels were determined by enzyme-linked immunoadsorbent assay and CCR5 expression, as well as the percentage of apoptotic lymphocytes, on a FACScan flow cytometer. After staining with different antibodies, the cells were subjected to three-colour flow cytometric analysis. Data analysis was performed using winmdi 2.5 software. The serum RANTES level and percentages of CCR5/CD4 and CCR5/CD8 T lymphocytes in CR, AR and SE were lower than that in the control group (P <0.05). The level of CD4 and CD8 T lymphocytes in early apoptosis was higher in AR patients than in CR, SE or C groups (P <0.05). In the case of late apoptosis, the percentage of apoptotic/necrotic cells was higher in the CR than AR, SE or C groups (P <0.05). The RANTES serum levels and the percentage of peripheral CCR5 T lymphocytes would not indicate the renal allograft state. The increase of early apoptotic T lymphocytes could be a marker of AR process and could also indicate the initial step in reducing the cytotoxic T lymphocytes, thus favouring the graft evolution.     

供体凋亡淋巴细胞预输注对猪肝移植受体CD4+T、CD8+T、CD4+CD25+调节性T细胞的影响

背景：凋亡细胞能够主动调节机体的免疫功能,并能通过调节机体细胞免疫和体液免疫的途径诱导免疫耐受,但这些结果只在大鼠肝脏移植模型中证实。 目的：探讨通过⁶⁰Co γ射线体外处理后的供体淋巴细胞预输注诱导猪肝移植特异性免疫耐受的作用中,对淋巴细胞亚群的影响。 方法：建立非转流小型猪原位肝移植模型。将受体猪随机摸球法均分为2组：空白对照组,受体猪无特殊处理,行肝移植;淋巴细胞组：受体猪在肝移植前7 d经耳静脉注射⁶⁰Co γ射线处理过的5×10⁸个供体淋巴细胞。观察两组受体猪移植后的存活时间,移植后T淋巴细胞亚型CD4⁺T、CD8⁺T、CD4⁺CD25⁺Tr变化及病理。 结果与结论：移植后3 d,两组病理活检均呈急性中、重度排斥反应;移植后6 d,两组均呈急性重度排斥反应。移植后1,3,6 d CD4⁺T、CD8⁺T、CD4⁺CD25⁺Tr升降趋势,两组间差异无显著意义(P > 0.05)。提示,⁶⁰Co γ射线体外处理过的淋巴细胞预输注未能够诱导猪同种异体肝移植特异性免疫耐受,未能引起T淋巴细胞亚群变化有关。     

Concomitant augmentation of CD4+ CD29+ helper inducer and diminution of CD4+ CD45RA+ suppressor inducer subset in patients infected with human T cell lymphotropic virus types I or II.

To examine the immunomodulatory effects of HTLV infection, lymphocyte subset analysis was performed on patients infected with human T cell lymphotropic virus type-I (HTLV-I, n = 6) or -II (HTLV-II, n = 12) and on normal blood donors (n = 16). The percentages of total B lymphocytes (CD19), natural killer (NK) cells (CD16), T lymphocytes and their subsets (CD2, CD3, CD4, CD5, CD7, CD8), and IL-2R (CD25) were found to be within the range found in normal donors. However, the expression of CD8+ HLA-DR+ increased significantly in patients with HTLV-I or HTLV-II infection (14.1 +/- 3.9% and 9.7 +/- 2.4% respectively; P less than 0.01) when compared with controls (3.2 +/- 1.1%). In addition, there was a significantly greater proportion of CD4+CD29+ T lymphocytes (29.3 +/- 6.1% and 31.1 +/- 9.0%; P less than 0.05) with concomitant diminution of CD4+CD45RA+ T lymphocytes (8.3 +/- 3.3% and 11.4 +/- 1.5%; P less than 0.01) in patients infected with HTLV-I or HTLV-II respectively, when compared with controls. The increased percentage of CD4+CD29+ subpopulations showed a direct correlation (rs = 0.86; P less than 0.001) with HTLV-specific antibody production. No difference in the CD8 population coexpressing CD29 and S6F1 (an epitope of LFA-1) were observed in the HTLV-infected group when compared with normal donors and functional analysis exhibited minimal cytotoxicity against lectin labelled heterologous target cells. Thus, the shift in the suppressor/cytotoxic to helper/inducer 'memory' CD4+ may be associated with immunoregulatory abnormalities often found in persons infected with HTLV-I or HTLV-II.     

Cytolytic function for pseudorabies virus-stimulated porcine CD4+ CD8dull+ lymphocytes

We previously observed that pseudorabies (PRV) virus-specific killing in vitro was mediated by CD6+ CD8+ lymphocytes. Also a high percentage of CD4+ lymphocytes, among these CD6+ CD8+ lymphocytes, was observed. The purpose of this study was, therefore, to further characterize the killing ability of PRV-stimulated CD4+ CD8+ lymphocytes. Peripheral blood mononuclear cells (PBMC) were isolated from blood of PRV-immune pigs and were stimulated in vitro with PRV. After 6 days, the frequency of CD4+ CD8+ lymphocytes in peripheral blood was determined by flow cytometry analyses. Lymphocytes were separated using a magnet-activated cell sorter or a FACSVantage SE, and the cytolytic activity of the isolated populations was determined. Flow cytometry analyses demonstrated that PRV stimulation of immune PBMC resulted in the occurrence of 26% +/- 4% CD4+ CD8dull+ lymphocytes. We further demonstrated that killing by PRV-stimulated PBMC was mediated by CD4+ CD8dull+ T lymphocytes and CD4- CD8+ T lymphocytes (classic cytolytic T lymphocytes and natural killer cells). The CD4+ CD8dull+ T lymphocytes showed major histocompatibility complex (MHC) II-restricted PRV-specific killing. The CD4- CD8+ T lymphocytes showed both PRV-specific and natural killing. The CD4+ CD8dull+ lymphocytes, which are unique in the pig, seemed to have a more heterogeneous function than was earlier demonstrated. In conclusion, we demonstrated that PRV-specific CD4+ CD8dull+ lymphocytes are able to kill PRV-infected target cells in a MHC II-restricted manner.     

Induction of CD95 ligand expression on T lymphocytes and B lymphocytes and its contribution to apoptosis of CD95-up-regulated CD4+ T lymphocytes in macaques by infection with a pathogenic simian/human immunodeficiency virus

Using an established SIV/HIV-C2/1-infected cynomolgus monkey model displaying stable CD4+ T cell depletion, the kinetics of apoptosis and the levels of expression of CD95 membrane-associated CD95L on lymphocytes were investigated to test the involvement of the CD95/CD95L system in CD4+ T lymphocyte loss in vivo. Rapid depletion of CD4+ T cells occurred up to 2 weeks after infection, with chronic CD4+ T lymphopenia thereafter. During the initial CD4+ T cell loss, which was accompanied by viraemia, about 90% of the peripheral CD4+ T cell subset underwent spontaneous apoptotic cell death during 24 h of culture. Increased expression of CD95 was observed on both CD4+ and CD8+ T cell subsets, with CD95 expression on CD8+ cells declining rapidly, but high CD95 expression being maintained on CD4+ cells. Since CD95L was expressed on CD8+ T cells, B cells and to a lesser extent on CD4+ T cells, this suggests that CD95-mediated apoptosis might be controlled in an autocrine/paracrine fashion.     

CD4+ CCR5+ and CD4+ CCR3+ lymphocyte subset and monocyte apoptosis in patients with acute visceral leishmaniasis

The potential involvement of apoptosis in the pathogenesis of visceral leishmaniasis (VL) was examined by studying spontaneous and Leishmania antigen (LAg)-induced apoptosis using cryopreserved peripheral blood mononuclear cells (PBMC) of Sicilian patients with VL. Results indicate that monocytes and T lymphocytes from acute VL patients show a significantly higher level of apoptosis compared with that observed in healed subjects. The percentage of apoptotic cells was higher in monocytes than in T lymphocytes. T cells involved in programmed cell death (PCD) were mainly of the CD4(+) phenotype. In particular, the T helper 1-type (Th1) subset, as evaluated by chemokine receptor-5 (CCR5) expression, is involved in this process. Cell death in Th1-type uses a CD95-mediated mechanism. Furthermore, Th1-type CCR5(+) cells are prone to cell suicide in an autocrine or paracrine way, as attested by enhanced expression of CD95L in acute VL patients. The reduction in Th1-type cells by apoptosis was confirmed by the decrease in interferon-gamma secretion. In conclusion, apoptosis of monocytes, CD4(+) and CD4(+) CCR5(+) T cells could be involved in the failure of cell mediated immunity that is responsible for severe immune-depression in VL.     

Changes in sensitivity of peripheral lymphocytes of autoimmune gld mice to FasL-mediated apoptosis reveal a mechanism for the preferential deletion of CD4-CD8-B220+ T cells

During thymic selection 'mis-selected' CD8(+) T cells exit to the periphery where they are deleted by a Fas/FasL-mediated mechanism, presumably as a result of activation by self-antigens. In the absence of functional FasL, as is the case in autoimmune gld mice, these 'mis-selected' T cells develop into unique Thy1(+)CD4(-)CD8(-) TCRalphabeta(+)B220(+) lymphocytes [abnormal double negative T (DN T) cells]. Using bioactive FasL-bearing vesicles [FasL vesicle preparation (FasL VP)], we were able to induce acute apoptosis in freshly isolated lymphocytes and to demonstrate that peripheral lymphocytes of gld mice become more sensitive to the FasL-mediated apoptosis as they age. Furthermore, flow cytometric analyses indicated that within this peripheral lymphocyte population, the abnormal DN T cells were preferentially eliminated. The exquisite sensitivity of these abnormal DN T cells is attributed to their increased membrane Fas expression with a concomitant reduction of cytosolic FLIP(L). Our data support the hypothesis that specific components of the Fas-mediated apoptotic pathway are modulated in favor of the elimination of auto-reactive T cells as well as those CD8(+) T cells that are 'mis-selected' in the thymus and escape to the periphery.