慢性乙型肝炎患者外周血CD45RA+,CD45RO+T淋巴细胞亚群的特点及临床意义

目的探讨慢性乙型肝炎患者外周血CD4 CD45RA ,CD4 CD45RO ,CD8 CD45RA 和CD8 CD45RO T淋巴细胞亚群的特点及其与肝病病情的关系.方法采集46例轻中度慢性乙型肝炎患者、58例重度慢性乙型肝炎患者和30例健康人的外周抗凝血,应用流式细胞技术三色荧光分析法对其外周血中CD4 CD45RA ,CD4 CD45RO ,CD8 CD45RA 和CD8 CD45RO T淋巴细胞亚群进行检测.结果轻中、重度慢性乙型肝炎患者与正常人相比,其外周血中CD4 ,CD8 T细胞均无明显改变;CD8 CD45RA T细胞均明显降低,CD8 CD45RO T细胞均明显增高,而CD4 CD45RA ,CD4 CD45RO T细胞均无明显改变;重度慢性乙型肝炎患者与轻中度慢性乙型肝炎患者相比,CD8 CD45RA T细胞明显降低(P<0.05),CD8 CD45RO T细胞明显升高(P<0.05).结论乙型肝炎慢性化过程中,CD8 CD45RO T细胞起重要作用且与慢性乙型肝炎患者病情的进展呈正相关;检测CD4 CD45RA ,CD4 CD45RO ,CD8 CD45RA 和CD8 CD45RO T淋巴细胞亚群比检测CD4 和CD8 T细胞亚群能使我们更加正确、充分、全面地了解慢性乙型肝炎的发病机制和预后,从而有效地指导临床治疗.     

慢性乙型肝炎患者外周血CD45RA+,CD45RO+T淋巴细胞亚群的特点及临床意义

目的 探讨慢性乙型肝炎患者外周血CD4＋CD45RA＋，CD4＋CD45RO＋，CD8＋CD45RA＋和CD8＋CD45RO＋T淋巴细胞亚群的特点及及其与肝病病情的关系．方法 采集46例轻中度慢性乙型肝炎患者、58例重度慢性乙型肝炎患者和30例健康人的外周抗凝血，应用流式细胞技术三色荧光分析法对其外周血中CD4＋CD45RA＋，CD4＋CD45RO＋，CD8＋CD45RA＋和CD8＋CD45RO＋T淋巴细胞亚群进行检测，结果轻中、重度慢性乙型肝楚患者与正常人相比．其外周血中CD4＋，CD8＋T细胞均无明显改变；CD8＋CD45RA＋T细胞均明显降低，CD8＋CD45RO＋T细胞均明显增高，而CD4＋CD45RA＋，CD4＋CD45RO＋T细胞均无明显改变；重度慢性乙型肝炎患者与轻中度慢性乙型肝炎患者相比，CD8＋CD45RA＋T细胞明显降低（P〈0．05），CD8＋CD45RO＋T细胞明显升高（P〈0．05）．结论乙型肝炎慢性化过程中，CD8＋CD45RO＋T细胞起重要作用且与慢性乙型肝炎患者病情的进展呈正相关；检测CD4＋CD45RA＋．CD4＋CD45RO＋．CD8＋CD45RA＋和CD8＋CD45RO＋T淋巴细胞亚群比检测CD4＋和CD8＋T细胞亚群能使我们更加正确、充分、全面地了解慢性乙型肝炎的发病机制和预后，从而有效地指导临床治疗。     

CD4~＋CD45~＋T细胞在反复上呼吸道感染患儿的表达及其与球蛋白的关系

目的：探讨反复上呼吸道感染（FURI）的患儿的CD4＋CD45＋T细胞两个亚群与球蛋白的特点。方法：收集现症及恢复期FURI患儿各26例,选取健康体检者18例作为对照;检测外周血CD4＋CD45RA＋及CD4＋CD45RO＋T细胞亚群的含量,并测定免疫球蛋白IgG、IgA及IgM的含量;且进行相关性分析。结果：健康组的CD4＋CD45RA＋T细胞显著高于恢复组,且恢复组高于现症组（P〈0.05）,但CD4＋CD45RO＋T细胞恰好相反,亦即现症组〉恢复组〉健康组（P〈0.05）;IgG及IgA的变化趋势为：现症组〈恢复组〈健康组,IgM的变化趋势与IgG及IgA相反;Pearson相关性分析显示CD4＋CD45RA＋及CD4＋CD45RO＋T细胞亚群呈显著负相关性（r=-0.791,P〈0.05）,CD4＋CD45RO＋T细胞与IgM呈正相关性,CD4＋CD45RA＋T细胞与IgG及IgA均呈显著正相关性（均为P〈0.01）。结论：反复上呼吸道感染患儿的CD4＋CD45RA＋/CD4＋CD45RO＋T细胞平衡向后者倾斜,且伴随体液免疫免疫失调,是该病的一个重要特征。     

关于CD45RA/CD45RO分子的功能研究

CD45RA和CD45RO是白细胞共同抗原的两种异型.CD45RA 和CD45RO T淋巴细胞在表型、功能等各方面均不同,了解其亚群的变化可能有助于了解许多疾病的发病机制、临床状态及预后.     

急性髓系白血病微小残留病灶检测方法的建立及其应用

目的 依据白血病相关免疫表型的抗原表达规律,建立急性髓系白血病微小残留病灶检测方法.方法 应用多参数流式细胞术对白血病细胞特征性抗原进行检测,所用荧光抗体涵盖了抗原跨系表达、抗原跨阶段表达,并注意抗原过度、过低或缺失表达以及光散射异常等情况,计算白血病细胞占全部检测细胞百分率,即MRD检测量.结果 23例患者抗原不同步表达CD33\CD13\CD15\CD34\CD45的抗体组合应用最多,占56.52％(13/23);CD33\CD13\CD15\CD117\CD45的抗体组合占21.74％(5/23).抗原跨系表达的抗体中CD33\CD13\CD19\CD34\CD45和CD33\CD13\CD7\CD34\CD45比例相对较少,分别为17.39％(4/23)和4.35％(1/23).生存期大于2 a的患者MRD数值明显低于生存期小于2 a的患者,差异具有统计学意义(P<0.01).结论 急性髓系白血病微小残留病灶检测是临床预后判断的重要参考指标.     

ROCK1 与CD40相互作用的质谱分析及在CD40信号通路中功能的初探

为鉴定和验证ROCK1和CD40的相互作用,探索ROCK1在CD40通路中可能发挥的作用,应用HD质谱分析ROCK1和CD40的相互作用,用免疫共沉淀实验验证其相互作用, 用ROCK1抑制剂研究ROCK1对CD40通路的影响. 结果ROCK1与CD40存在相互作用;并参与CD40介导的信号通路.说明ROCK1能与CD40发生相互作用,为CD40信号通路的研究提供了新的线索.     

基于CD/DVD光头的多层光存储系统

基于双光子吸收三维光存储技术和现有CD／DVD的高精度聚焦、循道伺服控制技术,搭建了一套与CD／DVD相兼容的信息存储系统．该系统通过DA输出选层信号控制音圈电机,实现多层读写．详细介绍了系统的工作原理．系统分为伺服和读写两个模块,伺服模块在跟踪光盘转动误差过程中提取光头的聚焦、循道伺服信号,同时为读写模块光头提供聚焦、循道激励信号．对双光头同步误差进行了测试,测试结果表明双光头激励信号具有一定的同步误差,基本符合系统正常运行时双光头同步误差的要求．     

表面等离子共振测定CD45-FITC抗体与蛋白A特异性结合常数

运用表面等离子共振(SPR)平台建立测定CD45-FITC抗体与蛋白A特异性结合常数的方法。当偶联蛋白A的传感芯片上流过CD45-FITC抗体溶液时,双通道SPR仪实时监测CD45-FITC抗体与蛋白A特异性结合过程,获得了该反应不同温度下的结合速率常数k_a、解离速率常数k_d和结合平衡常数K_a,以及该反应的活化能E_a和焓变△H。结果表明该特异性结合比较适宜的条件是:中性或微碱性、较高的离子强度和较高的温度。该方法操作步骤简单、快速、灵敏度高、消耗样品少,是研究分析免疫球蛋白IgG抗体和蛋白A等生物分子相互作用比较理想的方法之一。     

基于CD4512逻辑数据选择器设计

以CD4512八选一数据选择器作为核心元件,利用74LS161产生二进制的信号,当产生的信号与所需要的信号相同时,4 51 2输出高电平(1),若与要求不相符则输出低电平(0)。文章就是以实验设计逻辑数据选择器的过程讲述了74IS161同步加法计数器与八选一数据选择器。     

变频调速系统正弦信号部分产生的研究

本文阐述了以八阶低通滤波器MAX7480芯片为核心,利用锁相环CD4046,分频器CD4040以及其它元件组成的基本电路产生正弦波信号的原理。利用protel软件设计电路、在面包板上测试并且详细分析了设计思路,最终产生正弦波信号。     

Enhancement of T-cell responsiveness by the lymphocyte-specific tyrosine protein kinase p56lck.

Lymphocyte-specific tyrosine protein kinase p56lck is physically associated with CD4 and CD8 T-cell surface molecules, suggesting that it may transduce CD4/CD8-triggered tyrosine phosphorylation signals during antigen stimulation. Indeed, antibody-mediated aggregation of CD4 (to mimic interaction with its ligand, major histocompatibility complex (MHC) class II molecules), rapidly elevates the kinase activity of p56lck and is associated with marked changes in tyrosine protein phosphorylation. Genetic analyses suggest that the interaction of CD4/CD8 with p56lck results in a positive signal during antigen-induced T-cell activation. To evaluate directly the role of p56lck in T-cell activation, we introduced a constitutively activated form of Lck protein (tyrosine 505 to phenylalanine 505 mutant); in a CD4-negative, MHC-class II restricted mouse T-cell hybridoma. We report here that, as for transfection of CD4, expression of the Lck mutant enhanced T-lymphocyte responsiveness. This finding provides direct evidence that p56lck can positively regulate T-cell functions and that it mediates at least some of the effects of CD4 and CD8 on T-cell activation.     

CD3delta couples T-cell receptor signalling to ERK activation and thymocyte positive selection

Thymocytes from mice lacking the CD3delta chain of the T-cell receptor (TCR), unlike those of other CD3-deficient mice, progress from a CD4- CD8- double-negative to a CD4+ CD8+ double-positive stage. However, CD3delta-/- double-positive cells fail to undergo positive selection, by which double-positive cells differentiate into more mature thymocytes. Positive selection is also impaired in mice expressing inactive components of the Ras/mitogen activated protein (MAP) kinase signalling pathway. Here we show that CD3delta-/- thymocytes are defective in the induction of extracellular signal-regulated protein kinase (ERK) MAP kinases upon TCR engagement, whereas activation of other MAP kinases is unaffected. The requirement for CD3delta maps to its extracellular or transmembrane domains, or both, as expression of a tail-less CD3delta rescues both ERK activation and positive selection in CD3delta-/- mice. Furthermore, the defect correlates with severely impaired tyrosine phosphorylation of the linker protein LAT, and of the CD3zeta chain that is localized to membrane lipid rafts upon TCR engagement. Our data indicate that the blockade of positive selection of CD3delta-/- thymocytes may derive from defective tyrosine phosphorylation of CD3zeta in lipid rafts, resulting in impaired activation of the LAT/Ras/ERK pathway.     

Tyrosine phosphatase CD45 is essential for coupling T-cell antigen receptor to the phosphatidyl inositol pathway

Stimulation of T lymphocytes through their antigen receptor (T-cell receptor; TCR) results in the activation of a tyrosine kinase and the generation of phosphatidyl inositol (PtdIns)-derived second messengers. Several reports have indicated that CD45, a haematopoietic cell-specific surface glycoprotein with tyrosine phosphatase activity in its cytoplasmic domain, is important in lymphocyte activation. To examine the possibility that CD45 might influence proximal signal transduction events through the TCR, we have isolated a variant of the human T-cell leukaemic line, HPB-ALL, which fails to express this phosphatase. Unlike cells expressing CD45, stimulation of the TCR in the CD45-negative cell does not result in PtdIns-derived second messengers. Reconstitution of CD45 expression restored early signalling events through the TCR. To localize the site of CD45 action, the human muscarinic type 1 receptor, which also activates the PtdIns second messenger pathway, was transfected into the CD45-negative cell. Although stimulation of the TCR failed to generate PtdIns-derived second messengers, there was normal activity of the PtdIns pathway when human muscarinic receptor type 1 was stimulated, despite the absence of CD45. These data indicate that CD45 influences a cellular component that is essential for effective coupling of the TCR to the PtdIns second messenger pathway.     

Boron-based Drug Design for Cancer Therapy

1 Results Selective inhibition of protein tyrosine kinases is gaining importance as an effective therapeutic approach for the treatment of a wide range of human cancers.The epidermal growth factor receptor (EGFR) protein tyrosine kinase is one of the important kinases that play a fundamental role in cell growth signal pathways.We focused on the 4-anilinoquinazoline framework,which is observed in both compounds as a common structure.A boron atom has a vacant orbital and interconverts with ease between th...     

Intrathymic signalling in immature CD4+CD8+ thymocytes results in tyrosine phosphorylation of the T-cell receptor zeta chain

Thymic selection of the developing T-cell repertoire occurs in immature CD4+CD8+ double-positive thymocytes and is thought to be mediated by signals transduced by T-cell antigen receptor (TCR) molecules and possibly by CD4 and CD8 accessory molecules as well. It is not known, however, which signal-transduction mechanisms function in immature CD4+CD8+ thymocytes on engagement of TCR, CD4 or CD8 molecules. In mature T cells, CD4 and CD8 molecules are each associated with the src-like protein tyrosine kinase p56 lck and signals transduced by TCR and CD4 activate tyrosine kinases that phosphorylate TCR-zeta chains and other intracellular substrates. Consequently, we examined whether tyrosine kinases could be similarly activated in immature CD4+CD8+ thymocytes. Unexpectedly, we found that TCR-zeta chains from CD4+CD8+ thymocytes were already phosphorylated in vivo, and that dephosphorylation of this TCR subunit occurred on removal of CD4+CD8+ cells from their intrathymic environment. Rephosphorylation of TCR-zeta in cultured CD4+CD8+ thymocytes occurred rapidly in vitro, either in response to cross-linking of TCR, CD4 or CD8 by specific monoclonal antibodies, or on cell-cell contact. These observations indicate that tyrosine kinases are activated in vivo in immature CD4+CD8+ thymocytes undergoing thymic differentiation and selection. They also indicate that TCR, CD4 and CD8 molecules can function in CD4+CD8+ thymocytes as signalling molecules to activate tyrosine kinases and that phosphorylated TCR-zeta serves as a marker of these signalling events.     

Phosphorylation of GAP and GAP-associated proteins by transforming and mitogenic tyrosine kinases.

The critical pathways through which protein-tyrosine kinases induce cellular proliferation and malignant transformation are not well defined. As microinjection of antibodies against p21ras can block the biological effects of both normal and oncogenic tyrosine kinases, it is likely that they require functional p21ras to transmit their mitogenic signals. No biochemical link has been established, however, between tyrosine kinases and p21ras. We have identified a non-catalytic domain of cytoplasmic tyrosine kinases, SH2, that regulates the activity and specificity of the kinase domain. The presence of two adjacent SH2 domains in the p21ras GTPase-activating protein (GAP) indicates that GAP might interact directly with tyrosine kinases. Here we show that GAP, and two co-precipitating proteins of relative molecular masses 62,000 and 190,000 (p62 and p190) are phosphorylated on tyrosine in cells that have been transformed by cytoplasmic and receptor-like tyrosine kinases. The phosphorylation of these polypeptides correlates with transformation in cells expressing inducible forms of the v-src or v-fps encoded tyrosine kinases. Furthermore, GAP, p62 and p190 are also rapidly phosphorylated on tyrosine in fibroblasts stimulated with epidermal growth factor. Our results suggest a mechanism by which tyrosine kinases might modify p21ras function, and implicate GAP and its associated proteins as targets of both oncoproteins and normal growth factor receptors with tyrosine kinase activity. These data support the idea that SH2 sequences direct the interactions of cytoplasmic proteins involved in signal transduction.     

Profound block in thymocyte development in mice lacking p56lck.

The protein Lck (p56lck) has a relative molecular mass of 56,000 and belongs to the Src family of tyrosine kinases. It is expressed exclusively in lymphoid cells, predominantly in thymocytes and peripheral T cells. Lck associates specifically with the cytoplasmic domains of both CD4 and CD8 T-cell surface glycoproteins and interacts with the beta-chain of the interleukin-2 receptor, which implicates Lck activity in signal transduction during thymocyte ontogeny and activation of mature T cells. Here we generate an lck null mutation by homologous recombination in embryonic stem cells to evaluate the role of p56lck in T-cell development and activation. Lck-deficient mice show a pronounced thymic atrophy, with a dramatic reduction in the double-positive (CD4+CD8+) thymocyte population. Mature, single-positive thymocytes are not detectable in these mice and there are only very few peripheral T cells. These results illustrate the crucial role of this T-cell-specific tyrosine kinase in the thymocyte development.     

A motif in the alphabeta T-cell receptor controls positive selection by modulating ERK activity

Positive selection allows thymocytes that recognize an individual's own major histocompatibility complex (self-MHC) molecules to survive and differentiate, whereas negative selection removes overtly self-reactive thymocytes. Although both forms of thymic selection are mediated by the alphabeta T-cell receptor (TCR) and require self-MHC recognition, an important question is whether they are controlled by distinct signalling cascades. We have shown that mutation of an essential motif within the TCR alpha-chain-connecting peptide domain (alpha-CPM) profoundly affects positive but not negative selection. Using transgenic mice expressing a mutant alpha-CPM TCR we examined the contribution of several mitogen-activated protein kinase (MAPK) cascades to thymic selection. Here we show that in thymocytes expressing a mutant alpha-CPM receptor, a positively selecting peptide failed to activate the extracellular signal-regulated kinase (ERK), although other MAPK cascades were induced normally. The defect in ERK activation was associated with impaired recruitment of the activated tyrosine kinases Lck and ZAP-70, phosphorylated forms of the TCR component CD3zeta and the adaptor protein LAT to detergent-insoluble glycolipid-enriched microdomains (DIGs). Therefore, an intact DIG-associated signalosome is essential for sustained ERK activation, which leads to positive selection.