急性冠脉综合征患者辅助性T细胞亚群变化及意义

目的：探讨急性冠脉综合征(ACS)患者外周血辅助性T淋巴(Th)细胞亚群1和亚群2(TH1/Th2)的变化及意义。方法：64例ACS患者(其中急性心肌梗死(AMI)28例，不稳定心绞痛(UA)36例)、18例稳定型心绞痛(SA)、12例陈旧性心肌梗死(OMI)、21例胸痛综合征(CPS)患者和23例对照(C)外周血单个核细胞(PBMC)经植物血凝素(PHA)刺激培养后，应用酶联免疫吸附方法(ELISA)检测培养液上清中干扰素γ(IFN-γ)和白介素4(IL-4)水平。结果：ACS组培养液上清中IFN-γ水平及IFN-γ/IL-4比值明显高于SA组、OMI组、CPS组和C组，OMI组、SA组、CPS组与C组间无显著性差异；各组间IL-4水平无显著性差异。ACS患者培养液上清中IFN-γ水平随发病时间的延长而逐渐下降，但AMI患者IFN-γ高表达持续时间更长。结论：ACS患者存在Th1／Th2功能失衡，主要表现为Th1细胞功能亢进，可能是ACS的发病机制之一；AMI患者Th1／Th2功能失衡可能参与了AMI后自身免疫因素引起的心肌损伤和心室重塑过程。     

Th22及其效应因子生物学特性相关研究

新近发现一种CD4+T细胞新亚群Th22,表达CCR6、CCR10和CCR4,产生IL-22、TNF-α、IL-13,但不产生IFN-γ、IL-4、IL-17.不同于Th1、Th2、Th17,Th22有稳定的效应因子体系和独特的信号转导方式,其基因编码的蛋白及趋化因子参与组织重塑和新血管生成.Th22细胞主要参与皮肤的...     

Th22细胞的研究进展

Th22细胞是最近发现的CD4~+T细胞功能亚群,表达CCR6、CCR4和CCR10,产生IL-22和IL-13,但是不产生IFN-γ、IL-4和IL-17,是独立于Th1、Th2和Th17的细胞亚群.IL-6和TNF能够诱导初始CD4~+T细胞向Th22细胞分化.     

人外周血和扁桃体中Tfh细胞与其他Th细胞亚群之间的关系

目的比较观察人外周血和扁桃体Tfh细胞的表型以及与Th1、Th17、Th22细胞亚群之间的关系。方法分离正常人PBMC及扁桃体单个核细胞,利用anti-CD3+anti-CD28或PMA+ionomycin刺激后,采用ELISA和流式细胞术(FCM)检测其细胞因子的产生,分析Tfh与Th1、Th17、Th22细胞亚群之间的关系。结果与PBMC中CD4+T细胞不同,扁桃体CD4+T细胞高表达CXCR5和CD45RO,低表达CCR7和CD62L;与PBMC中CD4+T细胞相比,扁桃体CD4+T细胞IL-21和IL-17产生水平较高,IFN-γ产生水平较低,IL-22水平无显著差异;外周血和扁桃体CD4+T细胞中均存在一定比例的IL-21+IL-17+双阳性、IL-21+IL-22+双阳性、IL-21+IFN-γ+双阳性细胞,IL-21单阳性细胞在扁桃体CD4+T细胞中所占比例明显高于外周血;外周血和扁桃体CD4+CXCR5+细胞除表达IL-21外,还表达IL-17、IL-22和IFN-γ。结论扁桃体中存在较多数量的Tfh细胞,大多数Tfh细胞是不同于Th1、Th17和Th22的细胞亚群。     

Th22细胞在炎症免疫性疾病中作用的研究进展

Th22细胞是最近发现的CD4+T细胞功能亚群,表达CCR6、CCR4和CCR10,且分泌IL-22和TNF-α,不分泌IFN-γ、IL-4、IL-17,是独立于Th1、Th2和Th17的细胞亚群。Th22细胞主要参与皮肤的自稳调节和病理状态,可以调控皮肤的固有免疫反应、防御功能及表皮损伤后的修复功能,在炎症性皮肤病的病理生理机制中发挥重要作用。此外,Th22细胞也参与了炎症性肠病、类风湿关节炎、肝炎、试验性自身免疫性心肌炎等炎症免疫性疾病的病理过程。本文对Th22细胞其细胞因子IL-22在炎症免疫性疾病中作用的研究进展做一综述。     

急性心肌梗死大鼠心肌组织趋化因子及其受体mRNA表达的观察

目的:探讨急性心肌梗死大鼠心肌局部趋化因子和T细胞趋化因子受体的表达,揭示AMI后T细胞浸润心肌组织的机制。方法:结扎冠脉左前降支建立AMI大鼠模型,用半定量RT-PCR方法分析心肌梗死区和非梗死区趋化因子的表达,包括γ干扰素诱导的单核因子(MIG),正常T细胞表达和分泌、活化时表达下降的因子(RANTES),巨噬细胞炎性蛋白-1α(MIP-1α),以及T细胞趋化因子受体的表达(包括CXCR3、CCR3、CCR5)。HE染色切片进行心肌梗死区和非梗死区淋巴细胞计数分析。结果:心肌梗死区和非梗死区趋化因子RANTES、MIP-1α、MIG的mRNA表达于术后3天开始升高,1周达峰值,然后开始下降,8周降至正常,而趋化因子受体的表达没有明显改变。AMI大鼠心脏梗死区和非梗死区均可见淋巴细胞浸润,梗死区1周达高峰(81.0±10.3vs2.6±1.1,P<0.05),非梗死区2周达高峰(19.0±8.0vs3.2±0.8,P<0.05)。RANTES和MIP-1α的表达与淋巴细胞浸润显著相关。结论:AMI后心肌局部趋化因子表达增高,可能是诱导T细胞浸润心肌组织的始动因子。     

子宫内膜异位症患者腹腔Th17细胞和Th17.1细胞检测及致炎活性分析

目的:鉴别和分离子宫内膜异位症患者腹腔冲洗液中的辅助性T细胞17(Th17)和辅助性T细胞17.1(Th17.1),并比较二者致炎活性。方法:收集中重型子宫内膜异位症患者的腹腔冲洗液,根据细胞表面趋化因子受体CC趋化因子受体6(CCR6)、CC趋化因子受体10 (CCR10)、CC趋化因子受体4(CCR4)、CXC趋化因子受体3(CXCR3)表达差异,利用流式细胞术鉴别不同的腹腔T细胞亚群,检测其白介素-17A(IL-17A)、干扰素-γ(IFN-γ)和T-bet转录因子(T-bet)的表达,同时结合实时定量逆转录聚合酶链反应检测RAR相关孤儿受体C (RORC)、肿瘤坏死因子-α(TNF-α)、粒细胞-巨噬细胞集落刺激因子(CSF2)、白介素1受体Ⅰ型(IL1R1)等基因的mRNA水平,鉴别Th17和Th17.1细胞,并比较其致炎活性的差异。结果:腹腔Th17细胞的表型为CD3⁺CD4⁺CCR6⁺CCR10^-CCR4⁺CXCR3^-/lo, Th17.1细...     

人初始和记忆CD4+T细胞信使RNA基因芯片检测结果分析

目的探讨健康成人外周血初始和记忆CD4~+T细胞静息状态下表面分子、趋化因子受体、细胞因子和转录因子mRNA表达的差异。方法抽取健康成年人外周血,分离PBMC,染色后流式分选出CD45RO-的初始和CD45RO+的记忆CD4~+T细胞,裂解细胞,进行mRNA表达谱芯片检测。结果相比初始T细胞,静息状态下记忆CD4~+T细胞表达高水平的表面分子CTLA-4、PD-1、FAS、CD25,趋化因子受体CCR4、CCR6、CXCR3、CXCR5,细胞因子IFN-γ、TNF-α、IL-17和转录因子T-bet、EOMES、STAT4、GATA3和RORγt,并表达低水平的表面分子CD62L、CCR7、ICAM-I、CD40L,细胞因子IL-1β和转录因子NF-κB。结论静息状态下,与初始CD4~+T细胞相比,记忆CD4~+T细胞高表达某些活化分子、细胞因子、转录因子mRNA,可能是记忆CD4~+T细胞发生快速免疫应答的关键因素。     

IFN-γ/IL-4细胞因子失衡与乙型病毒性肝炎宫内感染的关系

宫内感染是乙型肝炎传播的主要途径之一,机体内HBV清除与1型T辅助细胞/2型T辅助细胞(Th1/Th2)细胞因子有关,通常将γ干扰素(IFN-γ)产生水平作为Th1细胞功能的指标,以白细胞介素4(IL-4)产生水平作为Th2细胞功能的指标,因此,研究IFN-γ/IL-4型细胞因子对阻断乙型肝炎病毒宫内感染具有重要意义。本文就IFN-γ/IL-4细胞因子失衡与乙型肝炎病毒宫内感染关系作一探讨。     

系统性红斑狼疮病人外周血Th细胞和Tc细胞IFN-γ及IL-4表达水平与疾病的关系

目的：观察SLE病人体内Th1／Th2及Tc1／Tc2比例失衡及其与疾病的关系。方法：SLE病人（活动期病人15例，稳定期病人20例）及正常人20例外周全血经PMA（20ns／ml）和离子霉索（ionomyein，1μmol／L）刺激4小时后，采用四色流式细胞术检测CD3^＋CD8^＋T细胞（主要为Th细胞）及CD3^＋CD8^＋T细胞（Tc细胞）胞浆内IFN-γ及IL-4的表达水平。血清抗dsDNA抗体检测采用ELISA法、血清免疫球蛋白和尿蛋白含量测定采用免疫速率散射比浊法。结果：SEE活动期病人Th细胞中IFN-γ的阳性百分率明显低于稳定期病人及正常人，Tc细胞中IFN-γ的阳性百分率明显低于正常人；IL-4的阳性百分率各组问无明显差异。稳定期病人仅Tc细胞IFN-1的百分率明显低于正常人。血清抗dsDNA抗体（＋）病人Th和Tc细胞IFN-γ的阳性百分率明显低于抗dsDNA（-）病人；免疫球蛋白（IgG，IgA，IgM中任何一种）异常增高的病人Th及Tc细胞中虽IL-4的阳性百分率无变化，但平均荧光强度明显高于免疫球蛋白水平正常的病人。Th及Tc细胞中IFN-γ和IL-4的表达水平与尿蛋白含量无明显关系。结论：SLE病人体内存在1型及2型细胞因子的异常表达，并与疾病活动程度存在一定的关系，值得进行深入研究。     

Th1/Th2 Functional Imbalance After Acute Myocardial Infarction: Coronary Arterial Inflammation or Myocardial Inflammation

Objectives: The study clarified whether the T-helper (Th)1/Th2 imbalance existed only in coronary arterial inflammation or in both coronary arterial inflammation and myocardial inflammation and explored the significance of the imbalance of Th1/Th2 function after acute myocardial infarction (AMI). Background: There are two different inflammatory processes in patients with AMI: the coronary arterial inflammation that leads to the pathogenesis of AMI and the myocardial inflammation after AMI that leads to ventricular remodeling, which are positively and negatively regulated by Th1 and Th2 lymphocytes, respectively. Methods: Peripheral blood mononuclear cells from 33 AMI patients, 22 unstable angina (UA) patients and splenocytes from 35 AMI Wistar rats were collected. Cytokine-producing Th cells were ambulatorily monitored by 3-color flow cytometry. Interferon (IFN)-γ and interleukin (IL)-4 mRNA in the rat myocardium and chemokine receptors CCR3,CCR5 and CXCR3 mRNA on the surface of rat T-lymphocytes after AMI were measured by RT-PCR. Results: IFN-γ-producing T-cells significantly increased in patients with AMI and UA within 24 hours after the onset of symptom. The high ratio of IFN-γ-producing T-cells recovered 1 week after the onset in UA patients, while it could be examined 1 week and even 1 month after the onset in AMI patients. The up-regulation of Th1 cell function is consistent with bad heart function. There was no significant difference on the frequencies of IL-4-producing T-cells between each group. 1 week, 2 weeks and 1 month after AMI, IFN-γ mRNA increased in the myocardium of rats, but there was no significant change on global Th cell functions. Conclusions: Th1/Th2 functional imbalance exists in both coronary arterial inflammation and myocardial inflammation processes. The up-regulation of Th1 cell-functions may participate in the immune-mediated ventricular remodeling after AMI.     

Reduced production of both Th1 and Tc1 lymphocyte subsets in atopic dermatitis (AD)

An imbalance of interferon-gamma (IFN-γ)-bearing CD4⁺ T (Th1) cells in the pathogenesis of AD is well recognized; however, a possible role in AD for CD8⁺ T cells secreting Th1-like cytokines (Tc1) has not been properly addressed. In this study, two- and three-colour FACS analysis allowed us to discriminate the Th1 from the Tc1 subset. AD patients had half the number of IFN-γ-producing circulating T cells (P < 0.005; 13.6 ± 1.9% (mean ± s.d.)) compared with normal donors (25.0 ± 2.4%). Specifically, both Th1 (4.8 ± 0.7%) and Tc1 (8.1 ± 1.1%) cells in AD were decreased compared with Th1 (8.8 ± 0.8%) and Tc1 (15.0 ± 1.5%) cells in controls. Moreover, at the mRNA level, the ratios of IFN-γ/IL-4 and IFN-γ/IL-10 were lower in cells from AD patients compared with controls. In conclusion, the decrease of IFN-γ-producing T lymphocytes in AD is due to a reduction in both Th1 and Tc1 IFN-γ-secreting cells; this may not only contribute to the over-production of IgE, but also explain the high incidence of cutaneous infections observed in AD patients.     

Kinetics of IL-17- and interferon-γ-producing PLPp-specific CD4 T cells in EAE induced by coinjection of PLPp/IFA with pertussis toxin in SJL mice

Systemic administration of Pertussis toxin (PTX) abrogates T cell tolerance mediated by injection of neuroantigens in incomplete Freund's adjuvant (IFA) and causes experimental autoimmune encephalomyelitis (EAE). PTX concomitantly induces high frequencies of neuroantigen-specific IFN-γ- and IL-17-producing T cells. Both IL-17 and IFN-γ have been implicated as a key effector cytokines in the pathogenesis of EAE, possibly with different functions. We therefore investigated potential differences in the temporal and spatial kinetics of the PTX-induced neuroantigen-specific IFN-γ- and IL-17-producing T cell effector populations. IFN-γ- and IL-17-producing PLPp-specific T cells initially arose in comparable frequencies in the local draining lymph nodes (drLN) after immunization as measured by cytokine ELISPOT. High frequencies of both IFN-γ- and IL-17-producing T cells were present in the immune periphery before onset of EAE. The highest frequencies of PTX-induced IFN-γ- and IL-17-producing PLPp-specific cells coincided in the inflamed CNS during acute EAE. During recovery, both IFN-γ- and IL-17-producing PLPp-specific T cells simultaneously disappeared from the CNS, whereas high frequencies of these cells remained present in the immune periphery. The functional affinity of both IFN-γ- and IL-17-producing T cells did not change during EAE. Therefore, autoimmune pathology in this model did not correlate with specific PTX effects either on Th1 or Th17 cells regarding their kinetics and CNS migration.     

Soluble flagellin, FliC, induces an Ag-specific Th2 response, yet promotes T-bet-regulated Th1 clearance of Salmonella typhimurium infection

Clearance of disseminated Salmonella infection requires bacterial-specific Th1 cells and IFN-γ production, and Th1-promoting vaccines are likely to help control these infections. Consequently, vaccine design has focused on developing Th1-polarizing adjuvants or Ag that naturally induce Th1 responses. In this study, we show that, in mice, immunization with soluble, recombinant FliC protein flagellin (sFliC) induces Th2 responses as evidenced by Ag-specific GATA-3, IL-4 mRNA, and protein induction in CD62L(lo) CD4(+) T cells without associated IFN-γ production. Despite these Th2 features, sFliC immunization can enhance the development of protective Th1 immunity during subsequent Salmonella infection in an Ab-independent, T-cell-dependent manner. Salmonella infection in sFliC-immunized mice resulted in augmented Th1 responses, with greater bacterial clearance and increased numbers of IFN-γ-producing CD4(+) T cells, despite the early induction of Th2 features to sFliC. The augmented Th1 immunity after sFliC immunization was regulated by T-bet although T-bet is dispensable for primary responses to sFliC. These findings show that there can be flexibility in T-cell responses to some subunit vaccines. These vaccines may induce Th2-type immunity during primary immunization yet promote Th1-dependent responses during later infection. This suggests that designing Th1-inducing subunit vaccines may not always be necessary since this can occur naturally during subsequent infection.     

Anti-TNF therapy in patients with rheumatoid arthritis decreases Th1 and Th17 cell populations and expands IFN-γ-producing NK cell and regulatory T cell subsets

The aim of this work was to study the effect of anti-TNF treatment on CD4+ Th1, Th17 and regulatory T cells (Tregs), together with CD8+ T cells and NK cells from rheumatoid arthritis (RA) patients. For this purpose, 18 RA patients received adalimumab during 16 weeks and their peripheral blood lymphocytes were assessed by flow cytometry at the beginning and at the end of the study. We found that the proportion of Th17 cells was directly correlated with Th1 cells, but inversely correlated with IFN-γ-producing NK cells. A decrease was observed in Th1, Th17 cells and IFN-γ-producing CD8+ T cells by anti-TNF therapy. Conversely, the proportion of Tregs increased, as did the percentage of IFN-γ-producing NK cells. We postulate that a rise in IFN-γ production due to recovery of NK cells’ function, together with expanded Tregs, contribute to decrease the Th17 response in anti-TNF-treated RA patients.     

Interleukin-12 increases interleukin-4 production by established human ThO and Th2-like T cell clones

Interleukin (IL)-12 is a potent inducer of cell-mediated immunity: it favors the generation of interferon (IFN)-γ-producing T cells, increases IFN-γ production by T cells and natural killer cells and prevents the generation of a Th2 response in murine in vivo models. Nevertheless, the effects of IL-12 on an established Th2 response remain poorly documented. In the present paper, we analyzed the effect of IL-12 on the profile of lymphokines produced by established IL-4-producing Th0 and Th2-like human T cell clones (TCC) and by polyclonal T cells. We found that IL-12 (i) enhances, as previously reported, IFN-γ production by Th0 TCC and, to a smaller extent, by Th2-like TCC, (ii) increases the proliferation of Th0 and Th2-like TCC and (iii) unexpectedly, synergizes with T cell receptor-associated or nonspecific stimuli in increasing IL-4 production by these TCC. Thus, IL-12 potentiates the production of IFN-γ and also of IL-4 by established IL-4-producing TCC. Although IL-12 has been widely reported to induce a Th1 response and to prevent the development of a Th2 response in vivo, IL-12 may on the contrary potentiate an established Th2 response in humans.     

CCK-8对KLH免疫小鼠脾细胞Th1/Th2平衡的影响

目的： 探讨八肽胆囊收缩素（CCK-8）对Th1/Th2平衡的调节作用。方法: 给予BALB/c小鼠钥孔戚血蓝蛋白（KLH）免疫同时体内给予不同剂量的CCK-8,酶联免疫吸附试验(ELISA)检测其脾细胞培养上清中Th1型细胞因子γ-干扰素(IFN-γ)、白细胞介素-2（IL-2）和Th2型细胞因子白细胞介素-4（IL-4）、白细胞介素-5（IL-5）水平,逆转录聚合酶链式反应(RT-PCR)法检测脾细胞中IFN-γ、IL-2、IL-4、IL-5 mRNA表达;ELISA法检测血清中Th1型抗KLH抗体IgG2a和Th2型抗KLH抗体IgG1水平。结果: ①KLH免疫使小鼠脾细胞分泌Th1/Th2型细胞因子水平明显增高,mRNA表达增高,KLH免疫同时给予CCK-8可使脾细胞培养上清中IFN-γ、IL-2含量进一步增加和IFN-γ、IL-2mRNA表达增高,而使IL-4、IL-5含量降低,IL-4、IL-5 mRNA表达减低和降低IL-4/IFN-γ比值。②KLH免疫小鼠血清中IgG2a、IgG1发生不同程度增高,CCK-8可使其血清中IgG1水平减低而使IgG2a水平增高。结论: CCK-8可促进KLH免疫小鼠体内Th1反应,使Th2优势反应向Th1方向转变。     

Interleukin-15 induces interleukin-17 production by synovial T cell lines from patients with rheumatoid arthritis

IL-17-producing T cells (Th17 cells) are believed to contribute to local inflammation and joint damage in rheumatoid arthritis (RA). Limited data exist on Th17 cells located within the inflamed synovial tissue (ST) of patients with RA. Here, we aimed to generate polyclonal T cell lines (TCLs) from the RA ST and assess their cytokine production, including the effects of exogenous IL-15 on IL-17 production in vitro. For five patients with RA, polyclonal TCLs were established from ST obtained by joint surgery. Synovial TCLs were expanded and stimulated by anti-CD3/CD28 microbeads and exogenous cytokines. Cytokine production was assessed by culture supernatant analyses and intracellular flow cytometry, and TCLs were sorted based on their surface expression of CCR6. In addition to IL-17, we detected IL-6, IL-10, IFN-γ and TNF-α in the synovial TCL culture supernatants. Exogenous IL-15 increased the production of IL-17 as well as the other cytokines except IFN-γ. For IL-17, this effect was more pronounced after prolonged culture times. Intracellular flow cytometry confirmed the presence of IL-17+ and IL-17+ IFN-γ+ CD4+ T cells in the TCLs. IL-17+ and IL-17+ IFN-γ+ T cells were enriched in the CD4+ CCR6+ population. In conclusion, Th17 cells can be detected after polyclonal expansion and stimulation of RA synovial TCLs generated by joint surgery. The Th17 cells from the RA ST were enriched in the CD4+ CCR6+ population, and they were sensitive to exogenous IL-15. Th17 cells present within the synovial compartment may contribute to the RA pathogenesis and local joint damage.