支气管哮喘大鼠CD4+CD25+T细胞的变化及地塞米松干预作用的研究

目的 研究支气管哮喘(简称哮喘)大鼠模型支气管肺泡灌洗液(BALF)、血液、脾脏CD4+CD25+T细胞的变化,及地塞米松对CD4+CD25+T细胞的影响.方法 50只SD大鼠随机分为5组,空白对照(A)组,哮喘(B)组,地塞米松1(C)组、地塞米松2(D)组,地塞米松3(E)组.A组第l天给予腹腔注射生理盐水l ml,第15～21天每天给予生理盐水雾化.B、C、D、E组用卵蛋白建立哮喘大鼠模型,第1天,每只大鼠腹腔注射抗原l ml(卵蛋白1 mg+灭活百日咳杆菌9×106个+氢氧化铝干粉100 mg)混悬液,第15～21天给予1%的卵蛋白雾化30 min,C、D、E组于雾化后分别给予腹腔注射地塞米松0.2 mg/kg、1 mg/kg、2 mg/kg.采用流式细胞仪检测的方法 ,观察大鼠体内BALF、外周血、脾脏CD4+CD25+T细胞的变化及使用不同剂量地塞米松后对其的影响.结果 B组BALF、外周血、脾脏CD4+CD25+T细胞表达占CD4+T细胞的百分比分别是(42.21±5.62)%、(12.69±2.70)%、(11.15±1.05)%,A组结果 分别是(18.76±5.85)%、(6.21±1.73)%、(7.85±2.13)%.B组与A组比较,差异均具有统计学意义(P＜0.01,P＜0.01,P＜0.05);C组、D组、E组BALF中CD4+CD25+T细胞占CD4+T细胞的百分比表达分别是(10.49±4.03)%、(13.28±5.12)%、(7.51±5.39)%,显著低于A组和B组,(P＜0.05,P＜0.01);外周血中,C组(6.03±1.43)%、D组(4.88±0.95)%与A组(6.21±1.73)%比较,差异无统计学意义,E组(3.49±0.62)%与C组、A组比较,差异有统计学意义(P＜0.05).脾脏中,C组(7.25±1.82)%、D组(8.63±3.18)%与A组(7.85±2.13)%比较,差异无统计学意义,E组(3.38±1.37)%与C组、D组、A组比较,差异有统计学意义(P＜0.05).结论 CD4+CD25+T细胞在哮喘大鼠体内有明显的优势表达,可能是哮喘发病的机制之一.地塞米松可以抑制CD4+CD25+T细胞的表达.BALF内CD4+CD25+T细胞的变化与外周血和脾脏的变化具有一致性,监测外周血或脾脏CD4+CD25+T细胞变化可了解肺部情况.     

CD4+CD25+T细胞与支气管哮喘

本文就CD4^＋CD25^＋T细胞的调节机制，CD4^＋CD25^＋T细胞与变态反应的关系，以及CD4^＋CD25^＋T细胞在支气管哮喘发病机制中的作用作一综述。     

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.     

CD4+ CD25+ 调节性T细胞与自身免疫性甲状腺疾病

CD4+ CD25+ 调节性T细胞为新近发现的一群功能成熟的T细胞亚群.其特征性表达叉头盒蛋白3(Foxp3)分子,专职免疫无能和免疫抑制,在维持外周免疫耐受,防止自身免疫性疾病发病中起着极为关键的作用.CD4+ CD25+ 调节性T细胞在自身免疫性甲状腺疾病(AITD)发病中的作用引起了人们的关注.动物实验发现CD4+ CD25+ 调节性T细胞存在与否决定了实验动物是否发生实验性自身免疫性甲状腺炎(EAT)和Graves病.人体研究发现CD4+ CD25+ 调节性T细胞数目和功能异常与人AITD发生密切相关.这些研究结果提示,CD4+ CD25+ 调节性T细胞可能在AITD发病中起重要作用.     

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。     

高危后循环短暂性脑缺血发作患者CD4＋CD25＋调节性T细胞检测及意义

目的 探讨高危后循环短暂性脑缺血发作患者外周血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21例高危后循环短暂性脑缺血发作患者、19例眩晕综合征患者及20例健康成人外周血Treg占CD4+T细胞比例.结果 高危后循环短暂性脑缺血发作组外周血Treg/CD4+T细胞比例(5.66％±1.91％)显著低于眩晕综合征组(9.18％±2.26％)和健康成人组(9.21％±2.71％).结论 高危后循环短暂性脑缺血发作患者外周血Treg比例下降.Treg比例降低可能破坏了外周自身免疫耐受并参与了动脉粥样硬化的发生发展,可能是高危后循环短暂性脑缺血发作的发病机制之一.     

CD4+CXCR5+ T cells activate CD27+IgG+ B cells via IL-21 in patients with hepatitis C virus infection

BACKGROUND: Chronic hepatitis C virus (HCV) infection causes the skewing and activation of B cell subsets, but the characteristics of IgG+ B cells in patients with chronic hepa-titis C (CHC) infection have not been thoroughly elucidated. CD4+CXCR5+ follicularhelperT(Tfh)cells,viainterleukin (IL)-21 secretion, activate B cells. However, the role of CD4+CXCR5+T cellsintheactivationof IgG+ BcellsinCHCpatientsis not clear.
METHODS: The frequency of IgG+ B cells, including CD27?IgG+B and CD27+IgG+ B cells,the expression of the activation markers (CD86 and CD95) in IgG+ B cells, and the percentage of circu-lating CD4+CXCR5+ T cells were detected by flow cytometry in CHC patients (n=70) and healthy controls (n=25). The con-centrations of serum IL-21 were analyzed using ELISA. The role of CD4+CXCR5+ T cells in the activation of IgG+ B cells was investigated using a co-culture system.
RESULTS: A significantly lower proportion of CD27+IgG+ B cells with increased expression of CD86 and CD95 was observed in CHC patients.The expression of CD95 was negatively correlated with the percentage of CD27+IgG+ B cells, and it contributed to CD27+IgG+ B cell apoptosis. Circulating CD4+CXCR5+ T cells and serum IL-21 were significantly increased in CHC patients. Moreover, circulating CD4+CXCR5+ T cells from CHC patients induced higher expressions of CD86 and CD95 in CD27+IgG+B cells in a co-culture system; the blockade of the IL-21 decreased the expression levels of CD86 and CD95 in CD27+IgG+ B cells.
CONCLUSIONS: HCV infection increased the frequency of CD4+CXCR5+ T cells and decreased the frequency of CD27+IgG+B cells. CD4+CXCR5+ T cells activated CD27+IgG+ B cells via the secretion of IL-21.     

自身免疫性甲状腺疾病和CD4~+CD25~+调节性T细胞

自身免疫性甲状腺疾病(AITD)的发生及发展与CD4~+CD25~+调节性T细胞(Treg细胞)的数量和功能密切相关.动物实验证明Treg细胞可抑制AITD的发生.如果清除动物体内的该类细胞,可导致AITD发病或使原有的甲状腺疾病加重,Treg细胞通过抑制效应性T细胞的激活而发挥对AITD的影响作用.无论是胸腺还是外周,不同诱导体系来源的Treg细胞均对AITD有影响作用.     

Frequencies of circulating IL-17-producing CD4+CD161+ T cells and CD4+CD161+ T cells correlate with disease activity in rheumatoid arthritis

Abstract

Objective. Rheumatoid arthritis (RA) is a common autoimmune disease that is primarily driven by effector T cells, particularly Th17 cells, which are mainly contained within CD4+CD161+ T cells. Thus, we aimed to explore whether the frequencies of circulating IL-17-producing CD4+CD161+ T cells and CD4+CD161+ T cells were correlated with RA disease activity.

Methods. The surface phenotype and cytokine production of blood were analyzed by flow cytometry in 52 RA patients and 17 healthy controls. The disease activity was evaluated by the 28-joint disease activity score.

Results. The frequencies of circulating IL-17-producing CD4+CD161+ T cells and CD4+CD161+ T cells were increased in RA patients, and they were elevated in patients with active disease status compared to patients with low disease status. Furthermore, their frequencies were positively correlated with disease activity parameters. Receiver operating characteristic curve analysis revealed that IL-17-producing CD4+CD161+ T cell levels were able to distinguish disease activity with 60.7 % sensitivity and 87.5 % specificity, while CD4+CD161+ T cell levels showed 92.9 % sensitivity and 66.7 % specificity.

Conclusion. These results support the hypothesis that Th17 cells are involved in the pathogenesis of RA and suggest that circulating CD4+CD161+ T cells are a potential biomarker of RA disease activity.     

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。     

慢性HBV感染患者外周血CD4~+CD25~+调节性T细胞表达及其临床意义

目的探讨CD4+CD25+调节性T细胞与慢性HBV感染后不同临床转归和临床特点的相关性。方法在26例慢性乙型肝炎(CHB)患者、15例无症状HBsAg携带者(ASC)和11例肝炎肝硬化(LC)患者和16例正常对照者,分离外周血单个核细胞(PBMC),采用流式细胞仪检测CD4+CD25+调节性T细胞的表达水平。结果CHB组和ASC组的CD4+CD25+调节性T细胞占CD4+T细胞的百分率分别为4.40±2.76%和4.43±2.10%,均高于正常对照组(2.70±0.97%),差异显著(P0.01);CD4+CD25+调节性T细胞的表达水平与HBVDNA水平无相关性(r=0.018,P0.05);在HBeAg阳性与阴性组患者CD4+CD25+调节性T细胞的表达也无明显的差异(P0.05)。结论慢性HBV感染者外周血CD4+CD25+调节性T细胞水平升高,可能与HBV感染的慢性化有关。     

CD4+ T cells in lung tissue predict responsiveness to cyclosporin A in interstitial pneumonia

OBJECTIVE: A limited number of case studies have demonstrated the steroid-sparing and disease stabilization effects of cyclosporin A (CsA) combined with corticosteroid in patients with chronic interstitial pneumonia (IP). Although CsA is known to inhibit the proliferation and function of CD4+ T cells, it is not clear what type of IP is responsive to CsA administration. METHODOLOGY: In order to evaluate whether any lymphocyte subsets in alveolar tissue predict the responsiveness to CsA, morphometric analysis was performed on lung tissue obtained from six IP patients who were treated with a combination of CsA and steroid because the therapeutic effect of steroid alone had been limited. RESULTS: Cell densities of CD4+ T cells were significantly decreased in the alveolar tissues obtained from CsA responders (n = 3) compared to non-responders (n = 3) (120 +/- 86/mm2 vs 503 +/- 172/mm2, P=0.0495). CD4/CD8 ratios of CsA responders were also significantly lower than those of non-responders (0.315 +/- 0.070 vs 1.975 +/- 0.965, P=0.0463). However, cell densities of CD8+ T cell and areas of B cells aggregates were similar in both groups. CONCLUSIONS: The present observations, contrary to expectation, reveal that lower cell densities of CD4+ T cells and lower CD4/CD8 ratios are associated with responsiveness to CsA in combination therapy, suggesting that pharmacological actions other than suppression of CD4+ T cells explain the efficacy of CsA in patients with chronic IP.     

Mouse splenic CD4+ and CD8+ T cells undergo extensive apoptosis during a Plasmodium chabaudi chabaudi AS infection

The presence and phenotype of apoptotic lymphocytes was studied in spleen cell suspensions taken from CB6F1 mice infected with Plasmodium chabaudi chabaudi AS. High levels of apoptotic cells were found, associated with high parasitaemias and splenomegaly. This was also accompanied by expansion and disarray of spleen white pulp. Apoptosis levels lowered when parasitaemia was cleared, but were still higher than in normal mice. At this time, the spleen was diminishing in size and the white pulp was contracting and rearranging. When parasitaemia was patent, the cells most affected by apoptosis were CD4+ T cells followed by CD8+ T cells, and to a lesser extent B220+ B cells. When parasitaemia was cleared, CD8+ T cells and B220+ B cells returned to basal levels of apoptosis, while CD4+ T cells still had higher apoptosis levels than normal mice. A similar pattern of lymphocyte subpopulation apoptosis was found in infected BALB/c mice, despite the fact that, for this mouse model, it has been reported that B cells are the cells that are most affected by apoptosis. We consider that the high levels of apoptosis in CD4+ T cells when parasitaemias are still high are not easily explained by a normal mechanism of down regulation of the immune response.     

The immunosuppressive effects of CD4+CD25+ regulatory T cells on dendritic cells in patients with chronic hepatitis B

The characteristics and functions of CD4⁺CD25⁺ regulatory T cells (Tregs) have been well defined in murine and human systems. However, the interaction or crosstalk between CD4⁺CD25⁺ Tregs and dendritic cells (DCs) remains controversial. In this study, the effects of chronic hepatitis B (CHB) CD4⁺CD25⁺ Tregs on the maturation and function of monocyte‐derived DCs were examined. The results showed that CD4⁺CD25⁺ render the DCs inefficient as antigen‐presenting cells (APCs) despite prestimulation with CD40 ligand. This effect was marginally reverted by applying neutralizing antibodies (Abs) to IL‐10 and TGF‐β. There were an increased IL‐10 and TGF‐β secretion and reduced expression of costimulatory molecules in DC. Thus, in addition to a direct suppressor effect on CD4⁺T cells, CD4⁺CD25⁺ may modulate the immune response through DCs in CHB patients.     

CD4~+ CD25~+调节性T细胞与恶性腹水的相关研究进展

CD4+ CD25+调节性T细胞(CD4+ CD25+Treg细胞)是具有独特免疫调节功能的T细胞亚群,抑制免疫反应,在机体免疫稳态维持、肿瘤免疫及移植耐受等方面发挥重要的作用。近年来,调节性T细胞在肿瘤免疫及治疗的研究中受到越来越广泛的关注。现就调节性T细胞在恶性腹水方面的研究做一简要综述。     

慢性乙型肝炎患者外周血CD8+CD28+T淋巴细胞百分比的变化及其临床意义探讨

目的 探讨不同病程阶段的慢性乙型肝炎患者外周血CD8⁺CD28⁺T淋巴细胞百分比的变化,以及CD8⁺CD28⁺T淋巴细胞百分比变化与血清HBsAg水平的关系。方法 2018年4月~2018年8月我院诊治的慢性乙型肝炎患者88例,其中免疫耐受期20例,免疫清除期28例,非活动期20例,再活动期20例,另选择健康人20例。使用流式细胞术检测外周血CD8⁺CD28⁺T淋巴细胞百分比。结果 健康人与免疫耐受期患者外周血CD8⁺CD28⁺T淋巴细胞百分比分别为（26.1±3.5）%和（26.3±3.4）%,差异无统计学意义（P>0.05）;免疫清除期患者CD8⁺CD28⁺T淋巴细胞百分比为（40.1±4.7）%,显著高于健康人（P<0.05）;非活动期和再活动期患者外周血CD8⁺CD28⁺T淋巴细胞百分比分别为（20.3±2.2）%和（26.1±2.2）%,显著低于健康人（P<0.05）;外周血HBsAg低、中、高三组人群外周血CD8⁺CD28⁺T淋巴细胞百分比分别为（24.0±7.5）%、（28.4±8.9）%和（33.2±8.5）%,各组间差异有统计学意义（P<0.05）。结论 不同病程阶段的慢性乙型肝炎患者外周血CD8⁺CD28⁺T淋巴细胞百分比存在明显差异,可能与病毒长期刺激机体免疫系统,导致免疫系统功能失调有关,而这种失调可能参与了慢性乙型肝炎的发病过程。     

HIV/AIDS患者外周血CD4+ T细胞计数和淋巴细胞计数相关性的多中心研究

目的 对淋巴细胞计数预测CD4+ T细胞计数的准确性进行评价,为临床应用提供支持.方法 在全国23个分中心共筛查2 013例未接受抗病毒治疗的HIV/AIDS患者,经流式细胞术检测CD4+ T细胞计数,分析血常规当中的淋巴细胞计数和CD4+ T细胞计数之间的相关性,绘制ROC曲线判断淋巴细胞计数预测CD4+T细胞计数的准确性,并计算其敏感度、特异度、阳性预测值和阴性预测值.结果 2 013例HIV/AIDS患者的淋巴细胞计数和CD4+ T细胞计数分别为(1 600±670)×106/L和(244±148)×106/L,两者呈现显著正相关性(r=0.482,P＜0.000 1),以淋巴细胞计数预测CD4T细胞计数＜100×106/L、＜200×106/L和＜350×106/L的AUCROC分别为0.790(95％ CI0.761 ～0.818,P＜0.000 1),0.733(95％ CI0.710 ～0.755,P ＜0.000 1)和0.732 (95％ CI0.706 ～0.758,P ＜0.000 1).结论 在HIV/AIDS患者的临床诊治中,用淋巴细胞计数预测CD4+ T细胞计数有其实用价值,可以考虑在不具备CD4+ T细胞计数直接检测时作为替代指标用于监测疾病进展,也可以在获得CD4+ T细胞计数结果之前用来初步快速判断患者病情.     

CD4＋T淋巴细胞异常表达程序性细胞死亡因子4在不稳定型心绞痛患者PCI围术期的作用及意义

目的探讨CD4+T淋巴细胞异常表达程序性细胞死亡因子4(PDCD4)在不稳定型心绞痛患者经皮冠状动脉介入治疗(PCI)围术期的作用及意义。方法入选住院并行择期PCI的不稳定型心绞痛患者33例,同时选取29例只行冠状动脉造影检查而不行PCI的不稳定型心绞痛患者为对照组。分别于冠状动脉造影前或PCI术前、冠状动脉造影后或PCI术后18 h~24 h抽取新鲜外周血,免疫磁珠法分选出CD4+T淋巴细胞,荧光定量PCR检测PDCD4 mRNA表达,免疫印迹法检测PDCD4蛋白表达,酶联免疫吸附法检测血清肿瘤坏死因子α(TNF-α)浓度。结果与对照组相比,PCI组术后PDCD4 mRNA和蛋白表达明显升高(P<0.05)。PCI组术后血清TNF-α浓度较术前进一步升高(16.11±1.45 ng/L比7.60±0.75 ng/L;P<0.05),对照组无明显变化(P>0.05)。结论不稳定型心绞痛患者PCI术后CD4+T淋巴细胞PDCD4表达上调,从而增加PCI术后心肌的炎症反应。     

CD4~+CD25~+调节性T细胞在约氏疟原虫感染小鼠应答早期中的作用

目的探讨CD4+CD25+调节性T细胞在约氏疟原虫感染早期的作用及意义。方法用约氏疟原虫(致死型)感染DBA/2和BALB/c小鼠,计数红细胞感染率;在感染后第0d、3d、4d、5d和6d提取脾细胞应为,流式细胞术检测两种小鼠感染不同时间脾细胞悬液中CD4+T细胞和CD4+CD25+T细胞百分含量;ELISA法检测脾细胞培养上清IFNγ、IL10和TGFβ1水平。结果DBA/2小鼠的IFNγ水平在感染后第3d迅速升高后缓慢下降(P<0.01),CD4+CD25+T细胞数仅在感染后第5d出现有意义的升高(P<0.05),而IL10水平和CD4+T细胞数量都无明显改变。BALB/c小鼠的IFNγ水平在感染后第3d出现有意义的升高后迅速下降(P<0.01),CD4+CD25+T细胞数在感染后第3d开始明显升高,于感染后第5d出现下降(P<0.05),而IL10水平自感染后第3～6d持续维持高水平(P<0.05),CD4+T细胞数量于感染后第6天明显下降(P<0.05)。结论CD4+CD25+调节性T细胞在致死型约氏疟原虫感染BALB/c小鼠早期可能通过抑制性细胞因子IL10发挥免疫抑制作用。