肺结核患者IL-15和效应性CD8^＋T细胞的检测及意义

目的 通过检测肺结核患者IL-15水平及其效应性CD8^＋T细胞的变化,探讨IL-15在结核病免疫保护和结核病病情进展中的作用。方法 对98例肺结核患者进行血清IL-15水平及效应性CD8^＋T细胞检测,并结合痰涂片、病变程度等进行分析。结果 肺结核患者初治、复治组IL-15水平及效应性CD8^＋T细胞的百分比计数均较正常对照组下降（P〈0．05或P〈0．01）,复治组IL-15水平及效应性CD8^＋T细胞的百分比计数低于初治组（P值均〈0．01）;肺结核患者重度组IL-15水平及效应性CD8^＋T细胞百分比计数明显低于轻、中度组（P值均〈0．01）。痰菌阳性、痰菌阴性两组间IL-15水平及效应性CD8^＋T细胞百分比计数比较差异无统计学意义。结论 IL-15有望为结核病病情判断、内源性复发的评估提供一个新的指标。     

CD4+CD25+调节性T细胞对肺结核患者特异细胞免疫的调节作用

目的　探讨调节性T细胞对结核患者特异性细胞免疫的调节作用及其在结核病发生中的意义。方法 采用免疫磁珠从健康对照和结核患者外周血单个核细胞中分离CD4⁺CD25⁺调节性T细胞,观察其对结核患者外周血免疫反应,包括细胞增殖反应和细胞因子IFN-γ及IL-10分泌的影响。结果 体外消除CD4⁺CD25⁺调节性T细胞没有显著影响健康对照PBMC对BCG抗原的增殖反应,但可以显著增强结核患者PBMC对BCG抗原的细胞增殖反应、细胞因子IFN-γ及IL-10的分泌。分离的CD4⁺CD25⁺调节性T细胞能显著抑制结核患者CD4⁺CD25^-T细胞对BCG抗原及抗-CD3的细胞增殖和细胞因子IFN-γ分泌;CD4⁺CD25⁺调节性T细胞也抑制BCG刺激的CD4⁺CD25^-T细胞IL-10的分泌,但不影响抗-CD3刺激的IL-10分泌。结论 CD4⁺CD25⁺调节性T细胞可能通过抑制结核患者特异性细胞免疫应答促进肺结核病的发生发展。     

CD4~+CD25~+调节性T细胞对肺结核患者特异细胞免疫的调节作用

目的探讨调节性T细胞对结核患者特异性细胞免疫的调节作用及其在结核病发生中的意义。方法采用免疫磁珠从健康对照和结核患者外周血单个核细胞中分离CD4+CD25+调节性T细胞,观察其对结核患者外周血免疫反应,包括细胞增殖反应和细胞因子IFN-γ及IL-10分泌的影响。结果体外消除CD4+CD25+调节性T细胞没有显著影响健康对照PBMC对BCG抗原的增殖反应,但可以显著增强结核患者PBMC对BCG抗原的细胞增殖反应、细胞因子IFN-γ及IL-10的分泌。分离的CD4+CD25+调节性T细胞能显著抑制结核患者CD4+CD25-T细胞对BCG抗原及抗-CD3的细胞增殖和细胞因子IFN-γ分泌;CD4+CD25+调节性T细胞也抑制BCG刺激的CD4+CD25-T细胞IL-10的分泌,但不影响抗-CD3刺激的IL-10分泌。结论CD4+CD25+调节性T细胞可能通过抑制结核患者特异性细胞免疫应答促进肺结核病的发生发展。     

新发活动性肺结核患者治疗前后表达干扰素-γ的CD4+、CD8+T细胞百分比变化及意义

目的探讨新发活动性肺结核患者治疗2个月后表达干扰素-γ(IFN-γ)的CD~+_4、CD~+_8 T细胞百分比变化及意义。方法选取2016年8月至2018年8月我院收治的新发活动性肺结核病患者80例,采集患者治疗前和4联强化治疗2个月后全血,用流式细胞仪检测CD~+_4、CD~+_8 T细胞百分比及表达IFN-γ的CD~+_4、CD~+_8 T细胞百分比,用酶联免疫吸附法检测血清IFN-γ水平。结果:4联强化治疗2个月后,患者CD~+_4、CD~+_8 T细胞百分比均有下降趋势,但与治疗前比较差异无统计学意义(P0.05);表达IFN-γ的CD~+_4 T细胞百分比低于治疗前,差异有统计学意义(P0.05);表达IFN-γ的CD~+_8 T细胞百分比与治疗前比较差异无统计学意义(P0.05);血清IFN-γ水平低于治疗前,差异有统计学意义(P0.05)。结论新发活动性肺结核患者治疗2个月后表达IFN-γ的CD~+_4 T细胞百分比降低,监测表达IFN-γ的CD~+_4 T细胞对于评估抗结核疗效有一定的参考意义。     

T细胞亚群失衡及CD8+CD28+T细胞表达在AECOPD中的临床意义

目的探讨T细胞亚群失衡及CD~+_8CD~+_(28)T细胞表达在AECOPD中的临床意义。方法选取AECOPD住院患者40例作为研究对象,另选取40例健康非吸烟者作为对照组,两组人群在性别、年龄等一般资料方面无统计学差异。AECOPD患者予以抗感染、止咳祛痰等常规治疗2周。治疗前及治疗后1个月时均空腹抽血采用免疫比浊法定量检测CRP,流式细胞仪检测T淋巴细胞亚群;并抽取健康对照组空腹血进行对比。结果与健康对照组比较,COPD急性加重期患者外周血中CD~+_3、CD~+_4、CD~+_4/CD~+_8比值均显著下降,CD~+_8CD~+_(28)T淋巴细胞及CRP水平显著升高,二者存在正相关关系。在治疗后1个月AECOPD组患者外周血CD~+_8CD~+_(28)T淋巴细胞及CRP均较治疗前显著下降(P0.05),而CD~+_3、CD~+_4及CD~+_4/CD~+_8比值虽较治疗前升高,但差异无显著统计学意义(P0.05)。结论 AECOPD患者体内的T淋巴细胞数量减少且T细胞亚群失衡,患者体内T淋巴细胞免疫功能的恢复是一个慢性过程。CD~+_8CD~+_(28)T淋巴细胞参与了COPD急性加重的气道炎症反应,与CRP可作为反应慢性阻塞性肺疾病急性加重期气道炎症程度的敏感指标。     

慢性心力衰竭患者CD4+CD25+调节性T细胞检测及意义

目的:探讨慢性心力衰竭(CHF)患者外周血CD4 CD25 调节性T细胞(Treg)水平及意义。方法:采用流式细胞分析法,检测42例CHF患者(CHF组)和16例正常对照者(对照组)外周血CD4 CD25 Treg/CD4 T细胞比例。结果:CHF患者外周血CD4 CD25 Treg/CD4 T细胞比例[(12.2±3.8)%]显著低于对照组[(16.3±5.2)%]。CD4 CD25 Treg/CD4 T比例在缺血性心脏病者[(12.6±4.1)%]与非缺血性心脏病者(12.0±3.7%)间差异无统计学意义,但心功能NYHA分级Ⅲ~Ⅳ级者[(10.4±3.2)%]比例明显低于Ⅰ~Ⅱ级者[(13.3±3·8)%]。结论:CHF患者外周血Treg比例减少,且与心功能有一定关系。CD4 CD25 Treg比例降低可能打破了外周免疫耐受,参与了心力衰竭的发生发展。     

扩张型心肌病患者CD4~+CD25~+Foxp3~+ T细胞的检测及意义

目的:探讨扩张型心肌病(DCM)患者外周血CD4+CD25+Foxp3+T细胞的水平及意义。方法:采用流式细胞术检测DCM患者30例及健康对照组20例外周血CD4+CD25+T细胞和CD4+CD25+Foxp3+T细胞的比例。结果:DCM患者外周血CD4+CD25+T细胞占CD4+T细胞的比例为(8.53±1.64)%,显著低于健康对照组的(11.4±2.17)%,P0.01;DCM患者CD4+CD25+Foxp3+T细胞占CD4+T细胞比例为(0.99±0.54)%,显著低于健康对照组的(1.55±0.55)%,P0.01;且DCM患者心功能越差,CD4+CD25+Foxp3+T细胞占CD4+T细胞的比例越低。结论:DCM患者调节性T细胞比例的减少,可能打破了自身免疫耐受,发生了针对心肌抗原的自身免疫反应,参与了DCM的发病。     

CD8~+ T细胞在CD4~+ T细胞耗竭小鼠卡氏肺孢子虫肺炎的致病作用

[目的 ]验证参与卡氏肺孢子虫致病过程中 ,宿主炎性反应及其对肺功能的影响。 [方法 ]:将小鼠CD4+ T细胞耗竭或将小鼠CD4+ 和CD8+ T细胞都耗竭后 ,经气管接种卡氏肺孢子虫。观察在CD8+ T细胞缺如或存在的情况下 ,小鼠呼吸机能的改变和肺炎的程度。 [结果 ]耗竭CD4+ 和CD8+ T细胞后 ,小鼠虽发生PCP ,但呼吸频率无明显加快 ,肺组织损伤程度也较轻。相反 ,仅耗竭CD4+ T细胞保留CD8+ T细胞的小鼠的呼吸频率明显加快 ,肺内炎性细胞反应和肺组织损伤程度均较重。支气管肺泡灌洗液 (BALF)中的CD8+ T细胞和嗜中性白细胞的数量明显增多。 [结论 ]PCP的致病过程中 ,宿主的炎性细胞反应对肺功能有直接损伤作用 ,其中CD8+ T细胞似起主要作用。     

肺结核患者外周血CD+8 T细胞中IFN-γ 和miR-365a表达关系及意义

目的探究肺结核患者外周血CD_8~+ T细胞中γ干扰素(IFN-γ)和微小RNA(miR)-365a表达水平、相关性及临床意义。方法选取2017年3月～2017年6月青海省人民医院收治的肺结核患者53例为肺结核组,以同期进行体检的健康志愿者47例为对照组,磁珠法分离外周血中分离CD_8~+ T细胞,应用流式细胞术检测外周血CD_8~+ T细胞中IFN-γ表达水平,应用荧光定量PCR检测CD_8~+ T细胞中miR-365a表达水平,分析IFN-γ、miR-365a表达水平与临床参数的关系及两者表达的相关性。结果肺结核组外周血CD_8~+ T细胞中IFN-γ及miR-365a表达水平显著高于对照组,差异有统计学意义(P 0. 05)。抗结核治疗时间3个月、3～6个月、 6个月的CD_8~+ T细胞中IFN-γ、miR-365a表达水平比较,差异有统计学意义(P 0. 05),且随抗结核治疗时间的延长而降低(P 0. 05);治疗前痰抗酸染色涂片阳性的患者其CD_8~+ T细胞中IFN-γ、miR-365a表达水平显著高于痰抗酸染色涂片阴性的患者(P 0. 05)。肺结核患者CD_8~+ T细胞中IFN-γ与miR-365a表达水平呈显著正相关性(r=0. 786,P=0. 000)。结论肺结核患者外周血CD_8~+ T细胞中INF-γ、miR-365a表达明显升高,两者呈显著正相关,有望成为动态监测肺结核病情变化的参考指标。     

肺结核患者外周血CD_4~+、CD_8~+T细胞Blimp-1表达下降

目的研究活动性肺结核患者外周血单个核细胞(PBMCs)Blimp-1的表达及临床意义。方法采集31例活动期肺结核患者和45位健康对照组外周血,纯化PBMCs,用结核分枝杆菌ESAT-6和CFP-10混合性抗原肽库刺激,通过细胞表面标记和细胞内细胞因子染色技术,采用流式技术检测CD+4、CD+8T细胞Blimp-1的表达。结果与对照组比较,肺结核患者PBMCs中的CD+4、CD+8T细胞亚群分布出现显著性下降,且肺结核患者CD+4T细胞中Blimp-1的表达比例(%)下降(肺结核组89.5%(83.8%,95.7%),对照组94.5%(89.8%,98.7%),P0.05),且CD+4、CD+8T细胞中Blimp-1的表达量(平均荧光强度)也显著性下降(CD+4T细胞:肺结核组9.28(7.5,18.9),对照组15.4(11,25.4),P0.05);CD+8T细胞:肺结核组9.01(6.08,14.7),对照组14.2(9.53,23.1),P0.05)。结论活动期肺结核CD+4、CD+8T细胞群内Blimp-1的表达下降可能会使效应性和调节性T细胞的分化出现异常。Blimp-1可能参与结核病的疾病进程,这为研究结核病的诊断和治疗提供了线索。     

CD48在类风湿关节炎患者外周血CD8+T细胞上的表达及意义

目的探讨类风湿关节炎（RA）患者CD8^＋T细胞上CIM8的表达及意义。方法选择30例RA患者（RA组）和30名健康人（对照组），采用流式细胞仪测定两组外周血CD8^＋T细胞表面上CD48的平均荧光强度，分析RA患者CD48在CD8^＋T细胞上表达的临床意义。结果RA组CD48^＋CD8^＋T细胞表面平均荧光强度明显低于正常对照组，差异有统计学意义（P〈0．01）。而两组CD48^＋CD4^＋T细胞的表面平均荧光强度差异无统计学意义。结论RA患者CD8^＋T细胞上CD48的表达低于对照组，CD48的表达减低使抑制性T细胞减少可能在RA的发病过程中起作用。     

Induction of CD4+CD8+ double positive T cells and increase in CD5+ B cells in efferent lymph in sheep infected with Trypanosoma evansi

The effects of Trypanosoma evansi on efferent lymphocyte phenotypes draining from a lymph node primed with Pasteurella haemolytica vaccine were studied in sheep. The prefemoral efferent lymphatic ducts of the infected sheep along with those of two uninfected sheep were surgically cannulated. Lymph was collected and lymphocytes recovered from it analysed by two-colour indirect immunofluorescence staining and cytofluoremetry in a fluorescence activated cell analyser (FACSCAN). The study showed the appearance and persistence of T. evansi in the efferent lymph for a long period of time and the appearance of CD4⁺CD8⁺ (double positive, DP) T lymphocytes in the efferent lymph of infected animals. The infection also resulted in increases in CD5⁺ B cells in the prefemoral efferent lymph. In addition, there were decreases in the output of conventional B cells, CD5⁺ and CD4⁺ T cell subsets but large increases in CD8⁺ cells followed by terminal depletion of all cell subsets. In contrast, inoculation of sheep with pasteurella vaccine antigen alone produced little alterations in the proportions, but large increases in the numbers of all T cell subsets except that of CD8⁺ cells which also showed little variation; and there was a concurrent increase in the numbers and proportions of efferent B cells. In addition, the abnormal expression of DP and CD5⁺ B cells did not occur in the uninfected vaccinated sheep. It is concluded that these abnormal changes in the kinetics of efferent lymphocyte phenotypes are likely to play a role in the genesis of the generalized immunosuppression seen in trypanosome-infected hosts.     

Detection of intracellular IFN‐γ and IL‐4 cytokines in CD4+ and CD8+ T cells in the peripheral blood of dogs naturally infected with Leishmania infantum

Canine leishmaniosis (CanL) is a systemic zoonotic disease the clinical manifestations of which can range from self‐healing cutaneous lesions to disseminated visceral disease. Effective activation of cellular immunity is the cornerstone of resistance against Leishmania infantum in infected dogs. The aim of this cross‐sectional, controlled study was the intracellular detection of interleukin 4 (IL‐4) and interferon‐γ (IFN‐γ) in CD4+ and CD8+ lymphocytes in the peripheral blood of 40 dogs naturally infected with L. infantum by applying flow cytometry. The percentage of CD4+IL‐4+ and CD8+IL‐4+ lymphocytes (with or without immunostimulation) was low in the clinically healthy and subclinically infected dogs in contrast to clinically affected ones. In the same groups of dogs, the percentage of CD4+IFN‐γ+ and CD8+IFN‐γ+ T cells in their resting phase and following specific immunostimulation with Leishmania soluble antigen (LSA) was also low. CD4+IL‐4+ and CD8+IL‐4+ T cell percentage was higher in sick compared to clinically healthy and subclinically infected dogs, after immunostimulation. The corresponding figure of CD8+IL‐4+ cells in sick dogs after LSA immunostimulation was also increased thus underlining the important role these cells may play in humoral immunity and perhaps the progression of CanL.     

AIDS患者高效抗逆转录病毒联合治疗后外周血CD+8 CD+38T细胞与病毒载量同步变化

目的了解艾滋病(AIDS)患者高效抗逆转录病毒联合治疗(HAART)前后外周血CD+38抗原在CD+8T淋巴细胞上的表达情况.方法应用流式细胞仪采用双色荧光抗体检测技术检测CD+8 CD+38 T细胞;用罗式核酸扩增荧光定量聚合酶链反应(PCR)法检测血浆病毒载量(VL).结果 HAART后2周内CD+8 CD+38 T细胞数与VL开始同步下降,12周后83%AIDS患者的VL降至＜500拷贝/ml,同时CD+8 CD+38 T细胞计数与治疗前相比非常明显地降低(P＜0.001).而且63%的AIDS患者在血浆VL低于检测水平时,其CD+8 CD+38 T细胞数仍继续下降(与VL开始达到检测水平以下时相比,P＜0.001).结论 AIDS患者在HAART开始后,CD+8 CD+38 T细胞数与VL快速下降,在24周左右降至正常水平;并且CD+8 CD+38 T细胞数在VL达到检测不到时仍继续下降,提示在血浆VL低于检测水平时,CD+8 CD+38 T细胞数能够作为判断病毒是否复制的标记.     

Mechanisms regulating expansion of CD8+ T cells during HIV‐1 infection

Abnormal immune activation and expansion of CD8+ T cells, especially of memory and effector phenotypes, take place during HIV‐1 infection, and these abnormal features persist during administration of antiretroviral therapy (ART) to infected patients. The molecular mechanisms for CD8+ T‐cell expansion remain poorly characterized. In this article, we review the literature addressing features of CD8+ T‐cell immune pathology and present an integrated view on the mechanisms leading to abnormal CD8+ T‐cell expansion during HIV‐1 infection. The expression of molecules important for directing the homing of CD8+ T cells between the circulation and lymphoid tissues, in particular CCR5 and CXCR3, is increased in CD8+ T cells in circulation and in inflamed tissues during HIV‐1 infection; these disturbances in the homing capacity of CD8+ T cells have been linked to increased CD8+ T‐cell proliferation. The production of IL‐15, a cytokine responsible for physiological proliferation of CD8+ T cells, is increased in lymphoid tissues during HIV‐1 infection as result of microbial translocation and severe inflammation. IL‐15, and additional inflammatory cytokines, may lead to deregulated proliferation of CD8+ T cells and explain the accumulation of CD8+ T cells in circulation. The decreased capacity of CD8+ T cells to localize to gut‐associated lymphoid tissue also contributes to the accumulation of these cells in blood. Control of inflammation, through ART administration during primary HIV‐1 infection or therapies aimed at controlling inflammation during HIV‐1 infection, is pivotal to prevent abnormal expansion of CD8+ T cells during HIV‐1 infection.     

Polyfunctional Mycobacterium tuberculosis-specific effector memory CD4+ T cells at sites of pleural TB

Pleural tuberculosis (TB) is a common presentation of Mycobacterium tuberculosis (MTB) infection, and despite spontaneous resolution remains a strong risk factor for reactivation pulmonary TB in a majority of individuals. This study was undertaken to further understand the characteristics of immune cells at sites of pleural TB. A significant shift toward memory CD4+ T cells with an effector phenotype and away from na?ve CD4+ T cells in pleural fluid as compared to blood mononuclear cells was found. These data suggest that effector T cells are capable of migrating to sites of active TB infection and/or the differentiation to effector phenotype T cells in situ is highly amplified. Using multi-parameter flow cytometry analysis, a significant portion of MTB-specific CD4+ T cells in the pleural space were polyfunctional demonstrating two, three or four simultaneous functions including IFN-gamma, IL-2, TNF-alpha, and or MIP-1 alpha production. A greater proportion of these polyfunctional cells were of effector memory rather than central memory phenotype. The role of these polyfunctional MTB-specific CD4+ T cells at sites of pleural TB requires further study.