Antigen-specific activation and proliferation of CD4+ and CD8+ T lymphocytes from brucellosis patients

Salt-extractable antigen from Brucella melitensis 16M (RCM-BM) was used to evaluate the immune response from acute and chronic patients suffering from Brucella infections (in Mexico); their responses were compared with those of healthy controls. As a readout we used upregulation of CD69 (a well-established early activation marker for lymphocytes), lymphocyte proliferation by 3[H]thymidine or 5-bromo-2-deoxyuridine (BrdU) incorporation measured by liquid scintillation or flow cytometry, respectively, and production of gamma interferon (IFN gamma). We compared the antigen-specific response with the response induced by phytohaemagglutinin (PHA) as a positive control. There was no difference between acute patients and the healthy controls in the percentages of CD3+, CD4+ or CD8+ lymphocytes. However, we found that chronic patients had a significant (P < 0.05) increase in the CD8+ T cells, in line with previous studies. Antigen-specific responses to RCM-BM showed a significant (P < 0.05) upregulation of CD69 in both CD4+ and CD8+ T lymphocytes in acute brucellosis patients and in CD8+ T lymphocytes in chronic patients, indicating that both populations became activated by this antigen preparation. Moreover, lymphocyte proliferation from both acute and chronic patients in response to RCM-BM was highly significant (P < 0.001) when compared with healthy controls. However, there were no apparent differences between acute and chronic patients. Although the incorporation of BrdU showed similar results it provided additional information, since we demonstrated that both CD4+ and CD8+ T lymphocytes from acute and chronic patients proliferated equally well in response to RCM-BM. Similar results were observed with intracellular IFN gamma determination. As a whole, our data suggest an important role for both CD4+ and CD8+ T lymphocytes in Brucella infection in humans. As has been reported in mice, it is feasible that activated CD8+ T cells participate in protection against Brucella in humans through cytotoxicity or/and by the production of factors such as interferon and granulysin. The role of these cells should be carefully analysed to understand better their participation in human infection by Brucella.     

湖南省苗族健康人群外周血CD3+、CD4+、CD8+T淋巴细胞正常参考值研究

目的了解并建立湖南省苗族健康人群外周血CD3＋、CD4＋、CD8＋T淋巴细胞正常参考值。方法应用FACSCalibur流式细胞仪检测10-19岁、20-29岁、30-39岁、40-49岁、50-59岁5个年龄组共360名健康苗族人的CD3＋、CD4＋、CD8＋T淋巴细胞数值。结果10-59岁组苗族人群外周血CD3＋、CD4＋、CD8＋T淋巴细胞绝对数和CD4＋／CD8＋比值分别为（1426．71±462．92）个／μL、（787．54±258．36）个／μL、（589．86±242．91）个／μL和1．46±0．50;10～19岁年龄组的CD3＋、CD8＋T淋巴细胞计数均明显高于30岁以上年龄组（F分别为10．80、8．51,均P〈0．05）,CD4＋T淋巴细胞计数明显高于50-59岁组（F=7．03,P（0．05）;不同性别间上述指标无差异（t分别为0．98、0．80、0．88、0．16,均P〉0．05）。结论建立不同民族的CD3＋、CD4＋、CD8＋T淋巴细胞正常参考值有助于临床诊断。     

CD4+CD25+调节性T细胞与HIV感染关系的研究进展

CD4 CD25 调节性T细胞是一类具有特殊免疫调节功能的T细胞亚群。它能够抑制自身免疫病的发生,参与肿瘤免疫的调节,在感染和移植免疫中也发挥着重要作用。T细胞的这一亚群具有免疫无能和免疫抑制特性,新近发现它亦与艾滋病的发生、发展关系密切。文中就CD4 CD25 调节性T细胞与HIV感染的研究进展进行综述。     

CD4+CD25+T细胞与复发性流产

CD4+CD25+T细胞是一类具有免疫调节功能的细胞.正常情况下在体内发挥免疫抑制作用.该类细胞分泌白细胞介素10(IL-10)、转化生长因子β(TGF-β),表达CD25(IL-2Rα)、叉头蛋白(FOXP3蛋白)及细胞毒性T淋巴细胞抗原4(CTLA-4)分子等,通过细胞间直接接触或细胞因子间接方式广泛参与自身免疫耐受、肿瘤免疫、移植免疫.妊娠类似同种异体移植,妊娠时脱落人母体循环的胎儿抗原刺激CD4+CD25+T细胞的产生并在母胎界面形成免疫耐受微环境,防止胎儿受到母体排斥.CD4+CD25+T细胞数量或功能失调则会导致各种疾病的发生,如自身免疫病,移植物抗宿主病,流产等.在复发性流产患者体内CD4+CD25+T细胞数目及功能明显下降.实验表明增强CD4+CD25+T细胞数量或功能可以阻止移植物抗宿主病和治疗复发性流产.     

CD4~+ Notch1~+调节性T细胞和CD4~+ FoxP3~+调节性T细胞 在不明原因复发性流产中的表达

目的:探讨CD4+Notch1+调节性T细胞和CD4+FoxP3+调节性T细胞在不明原因复发性流产(URSA)中的表达。方法:流式细胞术分析复发性流产和正常妊娠动物模型中雌性小鼠CBA/J脾细胞CD4+CD25+Treg、CD4+FoxP3+Treg和CD4+Notch1+Treg的表达百分率变化。结果:流式细胞术分析(FACS)结果显示,相比于正常妊娠组,在复发性流产中CD4+CD25+Treg表达百分率减少、CD4+FoxP3+Treg表达百分率减少、CD4+Notch1+Treg表达百分率增加。结论:Notch1增加与FoxP3表达减少相关,CD4+Notch1+Treg的表达百分率增加可能是URSA的机制之一。     

CD4+T辅助细胞与常见自身免疫性疾病

CD4辅助性T细胞(T help cell/ Th)作为T细胞中的一个重要群体因分泌的细胞因子不同而被分为多个亚群,各个Th细胞亚群在自身免疫疾病发病中的作用在近十年中得到了广泛的研究及肯定.本文就Th细胞各亚群与自身免疫性疾病关系做一概述.     

Effects of occupational metallic mercury vapour exposure on suppressor-inducer (CD4+CD45RA+) T lymphocytes and CD57+CD16+ natural killer cells

Objectives: To examine the effects of metallic mercury vapour on the cellular and humoral immune system. Methods: We measured T lymphocyte and natural killer (NK) cell subpopulations, B lymphocytes, and serum immunoglobulins (i.e. IgG, IgA and IgM) together with total T (CD3+) lymphocytes and total lymphocytes in blood samples from 20 male, fluorescent-lamp makers (mercury workers) and the same number of gender-, age- and smoking-matched controls. Urinary concentrations of inorganic mercury (UHg) in the 20 workers ranged from 1.8 to 163.5 (mean 44.8) μg/l. They had been exposed to mercury vapour for 4 to 62 (mean 31) months. Results: Numbers of CD4+CD45RA+ (suppressor-inducer) T lymphocytes and total CD4+ T lymphocytes in the mercury workers were significantly smaller than those in the controls (paired-sample t-test, P < 0.01). The number of CD57+CD16+ NK cells was inversely correlated with UHg. Conclusion: It is suggested that numbers of CD4+CD45RA+ T lymphocytes and CD57+CD16+ NK cells are inversely affected by exposure to metallic mercury vapour in workers, with an average urinary inorganic mercury concentration of 45 μg/l being found. Received: 7 September 1999 / Accepted: 6 May 2000     

系统性硬皮病患者外周血CD4^＋T细胞中CD40L表达水平的研究

目的研究系统性硬皮病（SSc）患者外周血CD4^＋T细胞中CIM0配体（CIMOL）的表达水平。方法应用密度梯度离心法分离26例SSc患者（女16例,男10例）和25例正常对照者（女15例,男10例）的外周血单个核细胞,磁珠分选CD4^＋T细胞。RT—PCR检测CD4^＋T细胞中CD40L mRNA的表达水平;流式细胞术检测CD40L蛋白在CD4^＋T细胞中的表达水平。结果SSc女性患者CD4^＋T细胞中CD40L mRNA和CD40L蛋白的表达水平均显著高于正常女性对照组[5．61±1．86vs2．80±0．94,P〈0．01;（6．70±3．55）％vs（2．37±1．39）％,P〈0．05];SSc男性患者CD40L mRNA和CD40L蛋白的表达水平与正常男性对照组相比差异均无统计学意义[2．59±0．89vs1．92±0．56,P〉0．05;（2．06±1．09）％vs（2．13±0．87）％,P〉0．05]。结论女性SSc患者CD^＋T细胞中CD40L表达显著增高,而男性患者无明显改变,提示CD40L仅在女性SSc发病中起着重要的作用。     

子痫前期患者CD4~+CD25~+Foxp~(3+)调节性T细胞检测及意义

目的:研究CD4+CD25+Foxp3+调节性T细胞在子痫前期患者(PE)外周血中的比例变化,探讨其在子痫前期发病中意义。方法:采用细胞内染色的流式细胞术检测22例子痫前期患者外周血中CD4+CD25high和CD4+CD25+Foxp3+的表达,并与正常妊娠和正常育龄妇女进行比较。结果:子痫前期患者外周血单个核细胞(PBMCS)CD4+T细胞中CD25high细胞明显低于正常妊娠和正常育龄妇女,CD4+CD25+Foxp3+的表达显著降低,而且其CD4+CD25+Foxp3+的表达与平均动脉压呈负相关。结论:子痫前期患者CD4+CD25+Foxp3+的表达明显减少,表明调节性T细胞的降低可能打破了母胎之间的耐受,参与了子痫前期的病理进程。     

CD4+T细胞、CD8+T细胞、CD57+NK细胞和Vimentin在外阴硬化性苔藓中表达及临床意义

目的:探讨外阴硬化性苔藓中CD4+、CD8+T细胞及CD57+ NK细胞和Vimentin的表达.方法:采用免疫组化S-P法分别检测CD4+T细胞、CD8+T细胞、CD57+NK细胞和VIM阳性细胞在外阴正常皮肤(15例)、外阴硬化性苔藓早期(15例)及进展期(15例)病变中的浸润及表达.结果:CD4+T细胞在外阴硬化性苔藓早期中浸润明显高于正常皮肤,CD8+T细胞在外阴硬化性苔藓早期浅层及进展期深层病变组织中浸润均高于正常皮肤,CD57+ NK细胞在外阴硬化性苔藓早期中浸润明显高于正常皮肤,Vimentin标记的成纤维细胞在外阴硬化性苔藓早期浅层及进展期深层病变中表达明显高于正常皮肤,差异均有统计学意义(P＜0.05).结论:外阴硬化性苔藓中淋巴细胞的浸润与成纤维细胞增殖有一定相关性.     

外周血调节性CD_4~+ CD_(25)+ T细胞比例在体外受精与胚胎移植中的意义

目的:研究外周血调节性CD4+CD25+T(CD4+CD25+Tr)细胞的比例变化,探讨CD4+CD25+Tr细胞在妊娠免疫母胎耐受中的作用。方法:应用流式细胞术检测试管婴儿成功孕妇外周血中CD4+CD25+Tr细胞的比例变化。结果:试管婴儿成功孕妇外周血中CD4+CD25+Tr细胞比例较高,高于同孕期的正常妊娠组,差异有统计学意义(P<0.05)。结论:CD4+CD25+Tr细胞在体外受精与胚胎移植的发生发展中发挥一定作用。该研究将有助于阐明妊娠的本质,为探讨妊娠相关免疫异常疾病的发生机制及提高试管婴儿的成功率提供理论依据。     

Immunization with a soluble recombinant HIV protein entrapped in biodegradable microparticles induces HIV-specific CD8 cytotoxic T lymphocytes and CD4 Th1 cells

One of the major obstacles to the development of successful recombinant vaccines against human immunodeficiency virus (HIV) and other intracellular pathogens is the identification of a safe and effective vaccine delivery system for the induction of cell mediated immunity with soluble protein antigens. In this study it was demonstrated that immunization with a recombinant HIV envelope (env) protein entrapped in biodegradable poly(lactide-co-glycolide) (PLG) microparticles induced consistent HIV-specific CD4⁺ and CD8⁺ T-cell responses in mice. Major histocompatibility complex (MHC) class I-restricted cytotoxic T lymphocytes (CTL) responses were detected following a single systemic immunization with gp120 entrapped microparticles and when given by the intranasal (i.n.) route induced HIV-specific CD8⁺ CTL and secretory IgA. Furthermore immunization with gp120 entrapped in microparticles generated CD4⁺ T cells that secreted moderate to high levels of IFN-γ. Therefore, PLG microparticles are a safe and effective means of delivering antigen to the appropriate processing site for the generation of class I-restricted CTL, and are also capable of inducing Th1 cells.     

CD4+ T辅助细胞与常见自身免疫性疾病

CD4辅助性T细胞(T help cell/Th)作为T细胞中的一个重要群体因分泌的细胞因子不同而被分为多个亚群,各个Th细胞亚群在自身免疫疾病发病中的作用在近十年中得到了广泛的研究及肯定。本文就Th细胞各亚群与自身免疫性疾病关系做一概述。     

CD4+CD25+调节性T细胞在艾滋病病毒感染中的作用

CD4+CD25+调节性T细胞(Tregs)是一类具有特殊免疫调节功能的T细胞亚群,是Tregs的重要组成部分,参与多种感染性疾病的免疫应答,在病毒感染的慢性化、免疫病理、阻止自身免疫应答和维持机体免疫平衡等方面发挥重要的作用.此文就CD4+CD25+Tregs在HIV感染进程中的作用作了综述.     

CD4~+CD25~+调节性T细胞对狼疮样小鼠的干预性研究

目的分析CD4+CD25+调节性T细胞(regulatory T cells,Tregs)对狼疮样小鼠的干预效应,为系统性红斑狼疮(systemic lupus erythematosus,SLE)的免疫干预提供新的思路。方法采用磁活性标记的细胞分选方法(magnetic activated cell sorting,MACS)分选BALB/c小鼠CD4+CD25+T细胞,应用流式细胞术(flow cytometry,FCM)检测分选细胞纯度。将6~8w♀CB6F1小鼠随机分为3组,包括正常对照组、狼疮鼠模型组(亲代淋巴细胞免疫)、狼疮鼠干预组(亲代淋巴细胞免疫后2w,输入5×106Tregs/鼠)。分别于诱导后2、4、8、12w,采用ELISA法检测ANA和抗-dsD-NA抗体,并观察肾标本的病理学改变。结果BALB/c小鼠脾脏单个核细胞经MACS分选后,CD4+CD25+T细胞纯度达98.5%。经亲代淋巴细胞输注后,CB6F1小鼠出现体重降低、皮肤干涩;ANA和抗ds-DNA抗体上升;以及狼疮肾炎的病理学改变。而输入Tregs对已出现自身抗体的狼疮鼠有明显的逆转效应,CB6F1小鼠于输注后抗-dsDNA即...     

Development of HSV-specific CD4+ Th1 responses and CD8+ cytotoxic T lymphocytes with antiviral activity by vaccination with the HSV-2 mutant ICP10DeltaPK

A growth compromised herpes simplex virus type 2 (HSV-2) mutant which is deleted in the PK domain of the large subunit of ribonucleotide reductase (ICP10DeltaPK) protects from HSV-2 challenge in the mouse and guinea pig cutaneous and vaginal models and reduces the incidence and frequency of recurrent disease (Vaccine (17) (1999) 1951; Vaccine (19) (2001) 1879). The present studies were designed to identify the immune responses induced by ICP10DeltaPK and define the component responsible for protective activity. We found that ICP10DeltaPK elicits a predominant HSV-specific T helper type 1 (Th1) response, as evidenced by: (1) higher levels of HSV-specific IgG2a (Th1) than IgG1 (Th2) isotypes and (2) higher numbers of CD4+ IFN-gamma than IL-10 secreting T cells in popliteal lymph nodes. This Th1 response pattern was associated with a significant increase in the levels of IL-12 produced by dendritic cells from ICP10DeltaPK than HSV-2 immunized animals. Lymph node cells (LNCs) from ICP10DeltaPK immunized mice had significantly higher levels of HSV-2 specific cytolytic activity than LNCs from mice immunized with HSV-2 and it was mediated by CD8+ T cells. CD8+ CTL were not seen in LNCs from HSV-2 immunized mice. In adoptive transfer experiments, CD8+ T cells and, to a lower extent, CD4+ T cells from ICP10DeltaPK immunized mice inhibited HSV-2 replication, suggesting that they are involved in the protective immunity induced by ICP10DeltaPK vaccination.     

CD4^＋CD25^＋调节性T细胞对哮喘大鼠气道炎症的影响

目的观察CD4^＋CD25^＋调节性T细胞（CD4^＋CD25^＋Treg）对哮喘大鼠气道炎症的影响。方法将卵白蛋白（OVA）免疫耐受大鼠CD4^＋CD25^＋Treg细胞过继转移给哮喘大鼠,然后观察支气管肺泡灌洗液（BALF）中细胞计数及分类,ELISA检测BALF中IL-4、IL-5和IFN-1及血清OVA特异性IgE含量,HE染色观察肺组织的病理改变。结果与哮喘组比较,过继转移CD4^＋CD25^＋Treg细胞后哮喘大鼠BALF中细胞总数、中性粒细胞和淋巴细胞百分率降低（P〈0．05）,嗜酸性粒细胞（Eos）百分率明显降低（P〈0．01）;BALF中IL4和IL-5含量明显降低,IFN-1含量明显升高,血清OVA特异性IgE含量明显降低（P〈0．05）;气道炎症明显减轻。结论过继转移OVA免疫耐受大鼠CD4^＋CD25^＋Treg细胞可以明显抑制哮喘的慢性气道炎症。     

CD4~+ CD25~+调节性T细胞与卵巢癌的相关性研究

目的:探讨卵巢癌患者病灶间质中及上皮内CD3+TILs、CD4+TILs、CD8+TILs、Treg的分布特点。方法:采用免疫组织化学方法检测40例卵巢癌患者病灶间质中及上皮内CD3+TILs、CD4+TILs、CD8+TILs、Treg的表达情况及10例正常卵巢组织中CD3+T细胞、CD4+T细胞、CD8+T细胞、Treg的表达情况。结果:卵巢癌组织中CD3+T细胞、CD4+T细胞、CD8+T细胞数量与正常卵巢组织中相比显著增加,且卵巢癌组织上皮内Treg数量增加。结论:卵巢癌局部微环境中机体对肿瘤抗原可以产生免疫反应,但Treg的存在而抑制了肿瘤的特异性免疫。