活化条件对细胞内免疫染色IFNγ和IL-4法检测人外周血Th1和Th2细胞的影响

目的：研究细胞活化条件对Ｔｈ１和Ｔｈ２细胞测定的影响。方法：采用酶联免疫斑点法（ｅｎｚｙｍｅｌｉｎｋｅｄｉｍｍｕｎｏｓｐｏｔａｓａｙ,ＥＬＩＳＰＯＴ）测定活化细胞胞浆内的细胞因子。ＩＦＮγ阳性者为Ｔｈ１细胞,ＩＬ４阳性者为Ｔｈ２细胞。结果：ｉｏｎｏｍｙｃｉｎ＋ＰＭＡ,ＰＨＡ＋ＰＭＡ或Ｃａ２＋载体＋ＰＭＡ刺激５ｈ,ＩＬ４阳性率均在１３％～１４％;ＩＦＮγ阳性细胞分别为（７３４±４７１）％、（７５６±４７０）％和（８９７±５６０）％,而刺激１６ｈ时,ＩＦＮγ阳性率分别变为（１１１９±４７２）％、（６８８±１７５）％和（５９４±２８４）％,有明显差异。结论：活化条件影响Ｔｈ细胞亚群的测定。     

成人外周血αβ T细胞和γδT细胞体外活化CD69表达的比较性研究

目的：γδＴ细胞具有细胞毒效应，探讨γδＴ细胞体外活化的规律将有助于这类细胞的临床应用。方法：将成人外周血分别置于ＰＨＡ（４０ｍｇ／Ｌ）、ＰＤＢ（２×１０－７ｍｏｌ／Ｌ）及ＰＨＡ（４０ｍｇ／Ｌ）加上ＰＤＢ（２×１０－７ｍｏｌ／Ｌ）的条件下进行全血培养，在培养２ｈ和６ｈ后分别用双色免疫标记流式细胞法对外周血Ｔ细胞的ＴＣＲαβ、ＴＣＲγδ和ＣＤ６９分子进行分析。结果：Ｔ细胞在受刺激后２ｈＣＤ６９分子即有所增高，但其增加的程度因Ｔ细胞亚群和刺激剂不同而有差别，ＰＨＡ和ＰＤＢ分别使αβＴ细胞ＣＤ６９表达百分率由对照组的４４８±１３６升高到８７９±１６７和７３３６±２９８；ＰＨＡ和ＰＤＢ分别使γδＴ细胞ＣＤ６９表达百分率由对照组的１３６５±１７４升高到３５８４±１４０和９９０３±０９７。与２ｈ的情况相比，αβＴ细胞在各种刺激剂作用６ｈ后ＣＤ６９表达均有进一步的增高，γδＴ细胞在ＰＨＡ作用６ｈ后ＣＤ６９表达继续上调。结论：γδＴ细胞对多克隆活化剂的反应比αβＴ细胞更早和更强。这一特征更有利于体外扩增γδΤ细胞并将其用于临床免疫治疗。     

哮喘患者支气管肺泡灌洗液中细胞分类及其T细胞亚群的改变

目的和方法：采用间接免疫荧光法和支气管肺泡灌洗液（ｂｒｏｎｃｈｉａｌａｌｖｅｏｌａｒｌａｖａｇｅｆｌｕｉｄ，ＢＡＬＦ）技术观察１２例过敏性哮喘患者和２３例健康人外周血和ＢＡＬＦ中的Ｔ淋巴细胞亚群变化：结果：与健康对照组比较，过敏性哮喘患者外周血的Ｔ淋巴细胞亚群无明显改变，但其ＢＡＬＦ中的ＣＤ４＋细胞显著增高（５４．９７±４１４）％ｖｓ（７９．７１±９．６３）％，Ｐ＜００５；ＣＤ４＋与ＣＤ８＋细胞的比值也显著增高（１６４±０．３２ｖｓ２．３２±０．８３，Ｐ＜００５）。此外，过敏性哮喘患者ＢＡＬＦ中肥大细胞和嗜酸细胞百分比（００９±０．０４）％和（３６２±１．０６）％明显高于健康对照组（００２±０．０１）％和（０．３９±０．３０）％，Ｐ＜００５和Ｐ＜００１。结论：ＣＤ４＋细胞在哮喘的气道炎症中发挥重要作用。     

呼吸道合胞病毒感染患儿免疫功能变化及PBL体外表达IL－2R的动态观察

探讨７２例确诊呼吸道合胞病毒（ＲＳＶ）特异性血清抗体及鼻咽分泌物脱落细胞中ＲＳＶ抗原阳性患儿急性期及恢复期Ｔ淋巴细胞亚群，血清ＩｇＧ、ＩｓＭ、ＩｇＡ及ＩＬ－２Ｒ的活性表达，动态观察了呼吸道合胞病毒感染患儿急性期、恢复期和正常对照组外周血淋巴细胞经ＰＨＡ刺激后，于不同时间（２４ｈ、４８ｈ、７２ｈ）细胞膜上ＩＬ－２Ｒ的活性表达。结果表明，抗体效价恢复期较急性期升高４～１２８倍，病例组急性期ＣＤ３、ＣＤ１６、Ｂ细胞升高，ＣＤ４／ＣＤ８的比值下降，ＩｇＧ、ＩｇＡ均降低，而ＩＬ－２Ｒ的活性表达呈下降趋势。此结果有助于探讨ＲＳＶ感染患儿的免疫紊乱发生机制。     

肺癌患者血清免疫抑制酸性蛋白及T细胞亚群联合检测的意义

本文对４４例肺癌术前患者血清免疫抑制酸性蛋白（ＩＡＰ）含量及外周血Ｔ淋巴细胞亚群进行了检测,与正常组比较。结果表明：肺癌患者血清ＩＡＰ含量显著增高（Ｐ＜００１）,Ｔ亚群ＣＤ３＋,ＣＤ４＋细胞均显著降低（Ｐ＜００１）,ＣＤ８＋细胞显著增高（Ｐ＜００５）,ＣＤ４＋／ＣＤ８＋细胞比值亦显著降低（Ｐ＜００１）;并发现患者血清ＩＡＰ含量的增高与其外周血Ｔ细胞亚群ＣＤ４＋／ＣＤ８＋细胞比值的降低呈明显负相关（ｒ＝ ０４２;Ｐ＜００１）。在患者中有肺外转移者血清ＩＡＰ含量明显高于未转移者（Ｐ＜００１）;其Ｔ细胞ＣＤ４＋／ＣＤ８＋比值亦明显低于未转移者（Ｐ＜００１）。提出联合检测对患者免疫水平评估和预后的判断更具有一定的意义。     

胸腺摘除对重症肌无力患者T细胞亚群变化的影响

采用免疫荧光染色技术，经流式细胞仪分析，观察了重症肌无力（ＭＧ）患者胸腺摘除前、后外周血Ｔ细胞亚群变化，并研究了淋巴细胞经植物血凝素（ＰＨＡ）激活后Ｔ淋巴细胞亚群的变化。结果显示：１．未经治疗的患者存在明显Ｔ细胞亚群异常，表现为ＣＤ３~＋、ＣＤ４~＋ＣＤ８~－和ＣＤ２９~＋ＣＤ４~＋百分率高，ＣＤ４~－ＣＤ８~＋和ＣＤ１６， ５６~＋百分率低下；２．胸腺摘除后，ＣＤ３~＋、ＣＤ４~＋ＣＤ８~－、ＣＤ２９~＋ＣＤ４~＋百分率降低，ＣＤ４~－ＣＤ８~＋百分率增高；３．ＰＨＡ对外周血中ＣＤ３~＋和ＣＤ２９~＋ＣＤ４~＋有明显下调作用；ＣＤ４~＋ＣＤ８~＋亚群在２４小时呈现上调，４８－７２小时呈现下调；４．ＰＨＡ作用７２小时，正常人ＣＤ４~＋ＣＤ８~－亚群无明显变化，对ＣＤ４~－ＣＤ８~＋亚群呈下调作用；对摘除胸腺的患者ＣＤ４~＋ＣＤ８~－亚群呈现明显下调，对ＣＤ４~－ＣＤ８~＋亚群则呈轻度下调作用。本研究结果表明：摘除胸腺后，ＣＤ４~＋ＣＤ８~－亚群减少，ＣＤ４~－ＣＤ８~＋亚群占优势。     

羧甲基茯苓多糖对HPBL分泌IL—2,TNF,IL—6,IFN—γ的调节作用

用ＣＭＰ培养外周血淋巴细胞（ＨＰＢＬ）２４、３６、４８、７２ｈ采样检测的ＩＬ－２、ＴＮＦ、ＩＬ－６、ＩＦＮ－γ效价分别可达１３．６±４．３，４１．９±２．０，１８３７．４±４６４．３，１０３７．９±２１１．０Ｕ／ｍｌ，分别比无ＣＭＰ的细胞培养对照组的效价高０．８，７．４，０．５，１０．９倍（Ｐ＜０．０１），说明ＣＭＰ具有ＩＬ－２、ＴＮＦ、ＩＬ－６、ＩＦＮ－γ的诱生剂功能。由ＣＭＰ预处理ＨＰＢＬ后经ＰＨＡ和／或ＣｏｎＡ促诱生组的ＩＬ－２、ＴＮＦ、ＩＬ－６、ＩＦＮ－γ效价分别比无ＣＭＰ的ＰＨＡ和／或ＣｏｎＡ刺激的相应常规诱生组高１．２～２．８，０．５～１．１、０．５～０．８、０．４～０．６倍（Ｐ＜０．０１），尤以ＣＭＰ＋ＰＨＡ＋ＣｏｎＡ促诱生细胞因子效果最佳（Ｐ＜０．０１），说明ＣＭＰ又具有ＩＬ－２、ＴＮＦ、ＩＬ－６、ＩＦＮ－γ促诱生效应。     

抗胸腺细胞球蛋白在体外刺激T细胞增殖和释放细胞因子

抗胸腺细胞球蛋白作为一种免疫抑制剂在预防器官和多植的排斥反应和治疗严重的再生障碍性贫血中有确切的作用。本工作在体外研究了抗胸腺细胞球蛋白（ＡＴＧ）／ＰＨＡ刺激人ＰＢＭＣ的增殖和释放细胞因子。结果为（１）＾３Ｈ－胸腺嘧啶掺入试验表明ＡＴＧ与ＰＨＡ一样具有强的促有丝分裂活性。（２）ＡＴＨ刺激的Ｔ细胞是以辅助Ｔ细胞为主（ＣＤ４／ＣＤ８〉１），在同样的条件下ＰＨＡ刺激Ｔ细胞ＣＤ４／ＣＤ８的比值小１。（３）     

ConA激活的小鼠T细胞CD69表达动力学的体内外研究

目的 为明确ＣＤ６９体内外表达动力学并进一步探讨其作用。方法 ＣｏｎＡ体内外刺激小鼠Ｔ细胞后,选取不同时间点,流式细胞仪观察Ｔ细胞ＣＥ６９表达率。结果ＣｏｎＡ刺激后２ｈＣＤ６９即有明显表达,６～８ｈ达到峰值,ＣＤ８＾＝与ＣＤ８＾＋Ｔ细胞相比无明显差异。体外条件对ＣＤ６９的表达有显著的影响。结论 结果提示ＣＤ８＾－和ＣＤ８＾＋Ｔ细胞的活化均需要ＣＤ６９的参与,Ｔ细胞可能在活化早期一过性高表达ＣＤ６９     

McAb共激活T细胞介导肝癌细胞前后的膜分子与TCRVβ基因的表达

Ｔ淋巴细胞活化是一个涉及多种膜表面分子和受体以及一系列相关多肽的复杂过程，为了使Ｔ细胞发挥更好的识别和杀伤癌细胞的功能，采用抗ＣＤ３、ＣＤ２８、ＣＤ８０（Ｂ７１）、ＣＤ２、ＣＤ５８ＭｃＡｂ分别刺激健康人ＰＢＬｓ后作用肝癌细胞，对作用前后ＰＢＬｓ用ＦＡＣＳ进行表型分析，结果发现：作用后ＣＤ３和ＣＤ８分子表达比作用前明显增高，而ＣＤ４分子无显著变化，同时基因家族采用ＲＴＰＣＲＳｏｕｔｈｅｒｎ印迹分析ＴＣＲＶβ基因１～２０亚家族表达水平与特征，健康人ＰＢＬｓ分别加入ＩＬ２、ＰＨＡ、抗ＣＤ３和ＣＤ３＋ＣＤ２８、ＣＤ２８＋ＣＤ８０、ＣＤ２＋ＣＤ５８作用肝癌细胞（ＢＥＬ７４０２）前表达水平平均约为５％，作用ＢＥＬ７４０２后表达水平约为１３％～２５％，其特征为Ｖβ７增高，这提示在癌抗原的参与下ＭｃＡｂ共刺激的Ｔ细胞活化，ＴＣＲ接受ＡＰＣ相应抗原的刺激，具有该ＴＣＲ的淋巴细胞迅速增殖而成为针对抗原的Ｔ细胞克隆，发挥其识别和杀伤癌细胞的作用，这将为肿瘤生物治疗的研究提供分子免疫学依据。     

T cell homeostasis: keeping useful T cells alive and live T cells useful

It has become clear that the regulation of T cell numbers is under peripheral homeostatic control. However, the rules for homeostasis vary with the T cells' differentiation state and the overall T cell number in the animal. Furthermore, homeostatic pressures can cause unexpected changes in T cell differentiation and function which might promote or dampen T cell reactivity. In this review, we focus on the role of peptide/MHC and cytokine interactions in regulating the size and composition of the T cell pool.     

Designer T cells by T cell receptor replacement

T cell receptor (TCR) gene transfer is a convenient method to produce antigen-specific T cells for adoptive therapy. However, the expression of two TCR in T cells could impair their function or cause unwanted effects by mixed TCR heterodimers. With five different TCR and four different T cells, either mouse or human, we show that some TCR are strong--in terms of cell surface expression--and replace weak TCR on the cell surface, resulting in exchange of antigen specificity. Two strong TCR are co-expressed. A mouse TCR replaces human TCR on human T cells. Even though it is still poorly understood why some TCRalpha/beta combinations are preferentially expressed on T cells, our data suggest that, in the future, designer T cells with exclusive tumor reactivity can be generated by T cell engineering.     

Inhibition of T lymphocyte heparanase by heparin prevents T cell migration and T cell-mediated immunity

Previously we reported that activated T lymphocytes express a heparanase enzyme that degrades the heparan sulfate moiety of the proteoglycan of the extracellular matrix (ECM). Expression of the heparanase enzyme was found to be associated with the ability of activated T lymphocytes to penetrate blood vessel walls and accumulate in target organs. We recently found that relatively low doses of heparin administered to mice or rats inhibited T cell-mediated immune reactions. In the present study we investigated the effects in vitro and in vivo of the heparanase inhibitor, heparin, on the expression of T lymphocyte heparanase and on the ability of T lymphocytes to mediate a delayed-type hypersensitivity (DTH) reaction. We found that heparanase was induced by immunizing mice with antigen in vivo or by activating T lymphocytes with concanavalin A in vitro. Relatively low doses of heparin administered once daily in vivo (5 micrograms) or present in vitro (0.1 microgram/ml) inhibited the expression of heparanase induced by immunization or by concanavalin A incubation. Higher or lower doses of heparin did not have these effects. The same doses of heparin that inhibited expression of heparanase also inhibited the ability of the lymph node cells to migrate to a site of antigen and adoptively produce a DTH reaction. These findings suggest that modulation of cell-mediated immune reactions may be achieved by relatively low doses of heparin which inhibit expression of T lymphocyte heparanase.     

人的活化T细胞出现的T_4~+T_6~+双标记现象

OKT系列单克隆抗体的产生,促进了人T细胞亚类的划分。最初的报道认为,人的外周血T细胞可分为OKT_4~+(T_4~+)和OKT_8~+(T_8~+)细胞两个亚类,分别占T细胞总数的55～65%和30～40%。最近有报道指出,人的末梢血中存在少量的T_4~+T_8~+双标记细胞,在体外活化的T细胞,可以暂时     

T cytotoxic 1 and T cytotoxic 2 CD8 T cells both inhibit IgE responses

BACKGROUND: It is well recognized that CD8 T cells inhibit IgE responses. In this study, we investigated the mechanism of CD8 T cell-mediated IgE suppression by comparing the capacity of T cytotoxic 1 (Tc1) and T cytotoxic 2 (Tc2) CD8 T cells to inhibit IgE responses to ovalbumin (OVA). METHODS: Tc1 and Tc2 CD8 T cells were generated from OVA(257-264)-specific Vbeta5.2 T cell receptor (TcR) transgenic mice by stimulation with anti-CD3 and anti-CD28 under Tc1 and Tc2 polarizing conditions. Tc1 and Tc2 Vbeta5.2 TcR CD8 T cells (10(6)) were adoptively transferred to syngeneic mice, and following immunization with 100 micro of OVA/alum, serum IgE antibodies were measured by passive cutaneous anaphylaxis and expressed as the highest dilution that gave a detectable skin response. RESULTS: Both Tc1 and Tc2 CD8 T cells from OT-I mice inhibited IgE. CONCLUSION: Both Tc1 and Tc2 CD8 T cells promote Th1 immunity and inhibit IgE responses. This process appears to be independent of CD8 T cell-derived IFN-gamma, as both Tc2 (IFN-gamma-) and Tc1 (IFN-gamma+) CD8 T cells inhibited IgE.     

T1-selective diffusion weighted fMRI at 1.5T

Apparent diffusion coefficients (ADC) of protons contributing to the functional signal can be determined from diffusion weighted functional magnetic resonance imaging (MRI) studies. An earlier study indicated that ADCs calculated from the functional signal of an activated primary sensorimotor cortex are large, and consistent with a CSF or intravascular contribution to the functional signal. We have added inversion recovery pulses to isotropic diffusion weighted imaging to null CSF protons selectively within the imaging slice, or to null the outer volume blood flowing into the imaging slice. With the use of gradient recalled diffusion weighted echo-planar imaging at low gradient b factors, and without the use of inversion pulses, the ADCs x 10(3) in mm2/s (+/- SD) from the functional signal were 6.81 +/- 1.19. These ADCs were significantly higher than resting primary sensorimotor cortex ADCs of 2.26 +/- 1.49, measured at the same b factors. When CSF nulling was applied, the functional signal ADCs remained high. Application of inflow nulling decreased the functional signal to such a small value, that ADCs estimated from these functional signals were not assessed. The results are consistent with an intravascular contribution to the functional signal and to its large ADC.     

REDIRECTING T LYMPHOCYTE SPECIFICITY USING T CELL RECEPTOR GENES

Redirecting T cells by transferring T cell receptor (TCR) genes from tumor-associated antigen (TAA)-reactive T cell clones into human peripheral blood lymphocytes (PBL) has therapeutic potential for the treatment of diseases, including cancer. T cell specificity can be altered using retroviruses encoding TCR &#102 and TCR &#103 chain genes, or chimeric immunoglobulin (cIg) genes containing signaling domains of CD3 &#145 or Fc &#108 RI- &#110. This review evaluates recent studies using TCRs and cIgs to redirect T cell specificity and discusses some of the technical and biological hurdles that need to be addressed before these approaches can be successfully used to treat patients.