溃疡性结肠炎患者外周血Th17/Treg免疫平衡的研究

目的研究溃疡性结肠炎（UC）患者外周血Th17细胞（CD4＋IL-17＋T细胞）与Treg细胞（CD4^＋CD25^＋T细胞）数量以及相关细胞因子的表达水平,分析Th17／Treg细胞免疫平衡在UC发病机制中的作用。方法收集UC患者（UC组）和健康体检者（对照组）的外周肝素抗凝静脉血,分离纯化T淋巴细胞。分别以PE-CD4与FTTC—CD25单抗,PE-CD4与FITC—IL-17的单抗作双色流式细胞术,分析溃疡性结肠炎患者外周血Th17、Treg细胞的百分率,ELISA法检测血清中相关细胞因子IL-17和TGF—β1的水平。结果与对照组比较,溃疡性结肠炎患者外周血Th17细胞百分率升高,Treg细胞的百分率显著下降,Th17／Treg比值升高（P〈0．05）,血清中细胞因子IL-17升高、TGF-β1降低,并且IL-17／TGF-β1比值升高。结论溃疡性结肠炎患者外周血存在细胞免疫功能失调;Th17／Treg细胞数量与免疫平衡状态发生改变,T淋巴细胞耐受机制的破坏可能与溃疡性结肠炎的发病机制有关。     

强直性脊柱炎患者外周血T细胞亚群的变化及意义

目的探讨强直性脊柱炎（AS）患者外周血中T细胞亚群的变化及其意义。方法采用流式细胞仪（FCM）检测外周血淋巴细胞表面CD3、CD4、CD8及其胞质内细胞因子IFN-γ和IL-4的表达。结果与正常对照组相比,As患者外周血Th细胞（CD;CD4＋）百分率无显著差异,Tc细胞（CD;CD;）明显升高（P〈0．05）;Th1细胞（CD3^＋CD4^＋IFN-γ^＋）百分率明显升高,Th2细胞（CD3^＋CD4^＋IL4^＋）无显著性差异;Te1细胞（CD3^＋CD8^＋IFN-γ^＋）百分率明显降低,T02细胞（CD3^＋CD8^＋IL4^＋）无显著性差异。结论AS患者外周血T细胞Th1／Th2、Tcl／Tc2亚群比例失衡,呈Th1、Tc2优势型。     

CD4~+T细胞亚群Th1/Th2和Th17/Treg在卵巢癌中的分布及意义

目的探讨卵巢癌患者外周血中CD4~+T细胞亚群Th1/Th2和Th17/Treg及相关细胞因子的分泌及其意义。方法30例卵巢癌患者为研究组,20例健康体检者为对照组,采用流式细胞术、酶联免疫吸附法(ELISA)、RT-PCR检测两组外周血中Th1/Th2和Th17/Treg CD4~+T细胞亚群相关细胞因子、转录因子表达情况并对比。结果与对照组比较,研究组外周血中Th1细胞亚群降低,其分泌的相关细胞因子γ-干扰素(IFN-γ)、白细胞介素(IL)-2及T盒子21转录因子(T-bet)表达均显著降低(P0. 05);Th2,Th17,Treg细胞亚群比例均升高,其各自分泌的相关细胞因子IL-4、IL-5、IL-13,IL-17α、IL-17f、IL-21、IL-22,IL-10、IL-35、转化生长因子(TGF)-β及转录因子GATA3,维甲酸孤儿核受体家族(ROR)γt,叉头样转录因子(FOXP) 3表达均显著提高(均P0. 05)。结论卵巢癌患者外周血中Th1细胞亚群受到了抑制,Th2、Th17、Treg细胞亚群都得到促进,提示这些CD4~+T细胞亚群可以作为判断卵巢癌疾病进展的重要指标。     

双膦酸盐治疗骨质疏松导致的Th17与Treg细胞因子分泌失调的效果

目的探讨双磷酸盐治疗骨质疏松后对辅助性T细胞(Th)17/Treg细胞因子分泌的影响。方法采用酶联免疫吸附(ELISA)检测双膦酸盐治疗后,骨质疏松患者血浆中Th1、Th2、Th17、Treg细胞因子分泌情况;采用RT-PCR检测Th17与Treg特异转录因子RORγt与FOXP3的变化。结果双膦酸盐治疗后,相比正常人群,骨质疏松患者血浆中Th1、Th2两种细胞亚群细胞因子分泌水平差异无统计学意义(P0.05),而Th17细胞因子分泌显著下降,Treg细胞因子分泌显著上升(P0.05);RT-PCR结果显示,Th17与Treg细胞的特异转录因子RORγt显著下降,FOXP3显著上升(P0.05)。结论双膦酸盐治疗骨质疏松患者对Th1与Th2细胞因子分泌没有显著影响,而Th17细胞因子分泌显著下降,Treg细胞因子分泌显著上升。     

血管活性肽对自发性高血压大鼠免疫细胞群及相关细胞因子的影响

目的探讨血管活性肽(VIP)对自发性高血压大鼠(SHR)辅助T细胞(Th)1/Th2和Th17/Treg免疫细胞群及相关细胞因子分泌能力的影响。方法选用SHR 10只一组,分选na6ve CD4~+T细胞,在Th1、Th2、Th17、Treg极化条件下培养,加入或者不加入VIP处理。同时分析用VIP处理与不用VIP处理SHR脾脏中Th1/Th2和Th17/Treg免疫细胞群比例变化。用流式细胞仪、酶联免疫吸附(ELISA)、RT-PCR等方法检测相关细胞因子、转录因子表达情况。结果与没有加入VIP相比,加入VIP后,SHR动脉血压降低,Th2、Treg细胞亚群都得到了促进,相关的细胞因子分泌都增加。而Th1、Th17细胞亚群都被抑制,相关的细胞因子分泌都明显下降(均P<0.05)。结论 VIP可以促进SHR Th2与Treg亚群形成,抑制Th1、Th17细胞亚群的分化。     

Th17细胞／调节性T细胞平衡与自身免疫性肝病

最新研究发现了与Th1和Th2细胞亚群不同的新的活化CD4^＋T细胞亚群——调节性T细胞（Treg细胞）和Th17细胞,两者在发育和功能上互补,Th17细胞／Treg细胞平衡在自身免疫性肝病的发生、发展中发挥重要作用。本文就Th17细胞／Treg细胞平衡在自身免疫性肝病中的作用作一综述。     

香烟烟雾提取物对CD4^＋T细胞向Th17细胞和调节性T细胞分化的影响

目的观察香烟烟雾提取物（CSE）对CD4^＋T细胞向Th17细胞和调节性T细胞分化的影响,为探索香烟烟雾暴露导致气道慢性炎症的机制提供实验依据。方法采用免疫磁珠法分离纯化健康志愿者外周血CD4^＋T细胞,以1×10^6／ml细胞密度接种于96孔培养板,分为以下8个组①空白对照组;②T细胞刺激剂组：加入T细胞刺激剂抗人CD3／28抗体微磁珠;③T细胞刺激剂CSE干预组：CD3／28＋2%CSE;④细胞因子组：CD3／28＋细胞因子;⑤细胞因子CSE干预组：CD3／28＋细胞因子＋2％CSE;⑥芳香烃受体（AHR）激动剂6-甲酰基吲哚[3,2-b]咔唑（FICZ）组CD3／28＋细胞因子＋FICZ;⑦AHR拮抗剂白藜芦醇组：CD3／28＋细胞因子＋2％CSE＋白藜芦醇;⑧溶剂对照组：CD3／28＋细胞因子＋DMSO。细胞因子为含TGF-β/IL-1β／IL-6／IL～23的混合细胞因子,以诱导Th17细胞和调节性T细胞分化。培养5d后,收集细胞,采用流式细胞术检测细胞表面分子CD4^＋CD25^＋Foxp3^＋细胞（Treg细胞）,以及胞内细胞因子IL-17＋的CD4^＋T细胞（Th17细胞）,计算各种刺激培养条件下,诱导生成的Th17及Treg细胞占CD4^＋T细胞的百分比。结果①CSE对Th17细胞分化的影响：空白对照组Th17细胞比例为（0．69±0．12）％,加入T细胞刺激剂后为（1．32±0．12）％,在此基础上加入CSE的干预组IL-17＋细胞比例升高为（2．17±0．24）％,（t=3．21,P〈0．01）;细胞因子组IL-17＋细胞比例为（1．35±0．08）％,而在此基础上加入CSE的干预组Th17细胞升高为（2．58±0．39）%（t=3．13,P〈0．01）。②CSE对Treg细胞分化的影响：在空白对照组中,Treg细胞占CD4^＋T细胞中的比例为（0．21±0．19）％,在加入T细胞刺激剂后,Treg细胞比例升高（3．59±0．37）％,加入细胞因子后,Treg细胞比例明显增加（5．85±0．76）％;在继续加入CSE干预后,Treg细胞比例明显减少（3．07±0．33）％（t=3．74,P〈0．01）;同样,T细胞刺激剂CSE干预组也出现Treg细胞比例减少（2．19±0．19）％,（t=2．71,P〈0．05）。③AHR活化对Treg细胞和Th17细胞分化的影响：AHR激动剂FICZ组的Treg细胞比例明显低于细胞因子组[（2．60±0．40）％,（5．85±0．76）％]（t=4．18,P〈0．01）,但Th17细胞比例升高[（2．86±0．43）％,（1．35±0．08）％]（t=3．65,P〈0．01）。AHR阻断剂白藜芦醇组中的Treg细胞比例和Th17细胞比例均低于细胞因子组,分别为[（0．33±0．14）％,（5．85±0．76）％],（t=7．71,P〈0．01）和[（0．42±0．07）％,（1．35±0．08）％],（t=8．87,P〈0．001）,并且和空白对照组接近,在细胞培养第5天通过7-AAD-AnnexinV对该组细胞检测并未发现细胞异常凋亡或死亡情况,结合该组细胞培养过程中的镜下形态推测该实验组CD4^＋T细胞的分化增殖受到白藜芦醇抑制。结论CSE可促进CD4^＋T细胞向Th17细胞分化,并抑制Treg细胞分化,这一过程可能通过AHR诱导。     

Th17细胞与肝脏疾病

辅助性T细胞（Th细胞）根据产生细胞因子和生物学功能分为Th1和Th2细胞。最近研究发现了一种与Th1和Th2细胞亚群不同的活化CD4^＋T细胞亚群-Th17细胞。TGF-8与IL-6或IL-21的协同作用,诱导Th17细胞分化。IL-12家族的IL-23在促进IL-17分泌、增强Th17细胞效应功能方面发挥重要作用,而RORγt是其特异性转录因子。分化成熟的Th17细胞可以分泌IL-17A、IL-17F、IL-21、IL-22、IL-6、TNF—α等多种细胞因子,在介导炎性反应（防御病原菌感染）、自身免疫性疾病、肿瘤、移植排斥反应等过程中发挥重要作用。Th17细胞也为研究肝脏疾病的发病机制及防治策略提供了新思路和方向。     

白细胞介素家族在炎症性肠病发病中的意义

炎症性肠病（IBD）包括溃疡性结肠炎（UC）和Crohn病（CD）。根据所分泌细胞因子和介导免疫功能的不同，CD4^＋T细胞可分为1型辅助性T细胞（Th1细胞）和Th2细胞两型．Th1细胞以分泌干扰素（IFN）-1、白细胞介素（IL）-12等为主，Th2细胞以分泌IL-4、IL-10、IL-13等为主。Th1／Th2细胞亚群失衡一直被认为是IBD的发病机制之一。CD主要是，Th1型疾病，而UC主要是Th2型疾病。研究IL家族有助于探讨IBD的发病机制以及IL在IBD治疗中的作用。本文将IL家族分为致炎细胞因子和抗炎细胞因子两大类。分别介绍其在IBD发病中的意义。     

Th17细胞及其相关疾病的研究进展

Th17细胞是一类独立于Th1、Th2的CD4^＋T细胞亚群,在其分化过程中需要白细胞介素（IL）-6和转化生长因子-β、转录因子ROR-γt及STAT3等参与,分泌IL-17、IL-22、IL-6等多种细胞因子,并参与多种炎症反应、自身免疫性疾病的发生发展,本文旨在阐述Th17细胞及其相关疾病的研究进展作一综述。     

Involvement of Th1Th17 Cell Subpopulations in the Immune Responses of Mothers Who Gave Birth to Children with Congenital Zika Syndrome (CZS)

High levels of T helper 17 cell (Th17)-related cytokines have been shown in acute Zika virus (ZIKV) infection. We hypothesized that the high levels of Th17-related cytokines, associated with a regulatory environment during pregnancy, create a favorable milieu for the differentiation of CD4+Th17 cells. We present data from a cross-sectional study on mothers who confirmed ZIKV infection by qRT-PCR and their children. We also recruited non-pregnant women infected with ZIKV in the same period. ZIKV infection occurred between 2015 and 2017. We collected samples for this study between 2018 and 2019, years after the initial infection. We highlight that, after in vitro stimulation with ZIKV CD4 megapool (ZIKV MP), we found a lower frequency of IL-17-producing CD4+ T cells (Th17), especially in the mothers, confirmed by the decrease in IL-17 production in the supernatant. However, a higher frequency of CD4+ IL-17+ IFN-γ+ T cells (Th1Th17) responding to the ZIKV MP was observed in the cells of the mothers and children but not in those of the non-pregnant women. Our data indicate that the priming of CD4 T cells of the Th1Th17 phenotype occurred preferentially in the mothers who gave birth to children with CZS and in the children.     

外周血Th17细胞及IL-17^＋FoxP3^＋T细胞在非小细胞肺癌患者的表达变化

目的：探讨辅助性T17（Th17）细胞及白细胞介素17（IL-17）＋叉头蛋白3（FoxP3）＋T细胞在非小细胞肺癌患者外周血中的表达及意义。方法：57例非小细胞肺癌患者作为研究对象．25名正常人作为对照。采用流式细胞术检测外周血Th17细胞、调节性T细胞（Treg细胞）的百分率以及IL-17＋FoxP3＋T细胞占Treg细胞的百分率。结果：Ⅳ期非小细胞肺癌患者外周血单核细胞（PBMC）中Th17细胞百分率高于Ⅰ-Ⅲ期患者及正常对照（均P〈0．01）。非小细胞肺癌患者PBMC中的Treg细胞百分率高于正常对照,并且IL-17＋FoxP3＋T细胞占Treg细胞的百分率高于正常对照（P〈0．05）。结论：Ⅳ期非小细胞肺癌患者中Th17细胞以及II-17＋FoxP3＋T细胞数量增加,Th17以及IL．17＋FoxP3＋T细胞可能参与了非小细胞肺癌的肿瘤远处转移过程．从而影响肿瘤进程。     

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.     

Tumor-specific Th17-polarized cells eradicate large established melanoma

CD4+ T cells can differentiate into multiple effector subsets, but the potential roles of these subsets in anti-tumor immunity have not been fully explored. Seeking to study the impact of CD4+ T cell polarization on tumor rejection in a model mimicking human disease, we generated a new MHC class II-restricted, T-cell receptor (TCR) transgenic mouse model in which CD4+ T cells recognize a novel epitope in tyrosinase-related protein 1 (TRP-1), an antigen expressed by normal melanocytes and B16 murine melanoma. Cells could be robustly polarized into Th0, Th1, and Th17 subtypes in vitro, as evidenced by cytokine, chemokine, and adhesion molecule profiles and by surface markers, suggesting the potential for differential effector function in vivo. Contrary to the current view that Th1 cells are most important in tumor rejection, we found that Th17-polarized cells better mediated destruction of advanced B16 melanoma. Their therapeutic effect was critically dependent on interferon-gamma (IFN-gamma) production, whereas depletion of interleukin (IL)-17A and IL-23 had little impact. Taken together, these data indicate that the appropriate in vitro polarization of effector CD4+ T cells is decisive for successful tumor eradication. This principle should be considered in designing clinical trials involving adoptive transfer-based immunotherapy of human malignancies.     

Increased CD4~+CD45RA~-FoxP3~(low) cells alter the balance between Treg and Th17 cells in colitis mice

AIM To investigate the role of regulatory T cell(Treg) subsets in the balance between Treg and T helper 17(Th17) cells in various tissues from mice with dextran sulfate sodium-induced colitis.METHODS T r e g c e l l s, T r e g c e l l s u b s e t s, T h 1 7 c e l l s, a n d CD4+CD25+FoxP 3+IL-17+ cells from the lamina propria of colon(LPC) and other ulcerative colitis(UC) mouse tissues were evaluated by flow cytometry. Forkhead box protein 3(FoxP 3), interleukin 17A(IL-17A), and RORC m RNA levels were assessed by real-time PCR, while interleukin-10(IL-10) and IL-17 A levels were detected with a Cytometric Beads Array.RESULTS In peripheral blood monocytes(PBMC), mesenteric lymphnode(MLN), lamina propria of jejunum(LPJ) and LPC from UC mice, Treg cell numbers were increased(P 0.05), and FoxP 3 and IL-10 mR NA levels were decreased. Th17 cell numbers were also increased in PBMC and LPC, as were IL-17 A levels in PBMC, LPJ, and serum. The number of FrI subset cells(CD4+CD45RA+FoxP 3low) was increased in the spleen, MLN, LPJ, and LPC. FrI I subset cells(CD4+CD45RA-Fox P3high) were decreased among PBMC, MLN, LPJ, and LPC, but the number of Fr III cells(CD4+CD45RA-FoxP 3low) and CD4+CD25+FoxP 3+IL-17A+ cells was increased. Fox P3 m RNA levels in CD4+CD45RA-Fox P3 low cells decreased in PBMC, MLN, LPJ, and LPC in UC mice, while IL-17 A and RORC mR NA increased. In UC mice the distribution of Treg, Th17 cells, CD4+CD45RA-FoxP 3high, and CD4+CD45RA-FoxP 3low cells was higher in LPC relative to other tissues.CONCLUSION Increased numbers of CD4+CD45RA-FoxP 3low cells may cause an imbalance between Treg and Th17 cells that is mainly localized to the LPC rather than secondary lymphoid tissues.     

Prostaglandin E2-dependent IL-23 production in aged murine dendritic cells

CD4+ T cells of the Th17 subtype are over-represented in the aged immune system. Dendritic cells (DC) play a critical role in naïve CD4+ T cell differentiation. However, expression of cytokines by aged DC that promote differentiation or survival of Th17 cells has not been extensively investigated. Using bone marrow-derived DC from C57BL/6 mice of different ages we compared cytokine production after DC activation by Toll-like receptor agonists for TLR4 and/or TLR7/8. DC-derived TNF-α and IL-12p70 production and expression of DC co-stimulatory molecules did not vary significantly by age indicating that TLR expression, function and signal transduction were intact in aged DC. There were relatively minor age-related changes in TGF-β and IL-6 which promote Th17 differentiation, but IL-23, a Th17-suvival cytokine, increased more than 40-fold across the lifespan. DC-derived prostaglandin E2 (PGE2) also increased with age and the up-regulation of IL-23 expression by aged DC was blocked by indomethacin that prevents PGE2 production, and by antagonists of PGE2 receptors. Exogenous PGE2 added to DC cultures further enhanced IL-23 production from aged but not young DCs. These data indicate that age-related changes in DC PGE2 production are necessary, but not sufficient to induce DC IL-23 production. Such changes may play a role in the expansion of Th17 cells in the aged immune system.     

Th1/Th2 cells in patients with multiple myeloma

Multiple myeloma (MM) is a lymphoproliferative disorder that is characterized by a proliferation of clonal B cells in various stages of maturation that then infiltrate the bone marrow. MM has been reported to accompany various T cell abnormalities including quantitative and functional defects of CD4+ and CD8+ T cells. Recently, immunotherapy such as dendritic cell therapy, vaccination therapy, and anti-tumor antibody therapy, has been attempted in patients with MM. To develop more effective immunotherapy for patients with MM, further studies are required to identify the immunological abnormalities, especially in T cells, associated with MM. The T helper 1 (Th1) and T helper 2 (Th2) cells are characterized by distinct cytokine production patterns. The Th1 cells produce interferon gamma and interleukin-2 (IL-2), and are involved in cell-mediated immunity. The Th2 cells produce IL-4 and promote humoral immunity by stimulating antibody production, particularly IgE responses. Furthermore, Th1 and Th2 cells have been found to cross-regulate each other's development. The Th1/Th2 combination has an important role in immune response to many disorders including infection, autoimmune diseases, and malignancies. In this review, we report a Th1/Th2 imbalance in cases of MM, and discuss the relationship between T cell abnormalities and the pathology of MM.     

先天性心脏病患者免疫因子Th17/Treg失衡与肺动脉高压的关系

目的:探讨患者外周血中Th17细胞、CD4+D25+FoxP3+调节性T细胞(Treg)的变化及意义。方法:75例先天性心脏病(CHD)合并肺动脉高压(PAH)患者,分为轻度肺动脉高压组(28例)、中度肺动脉高压组(32例)及重度肺动脉高压组(15例),20例健康体检者作为对照组。采用流式细胞分析法检测外周血中Th17细胞、Treg细胞占CD4+T细胞的比例,分析Th17细胞与Treg细胞的比例与肺动脉压的相关性。结果:Th17/CD4+T细胞比例、Treg/CD4+T细胞的比例和Th17/Treg细胞比值,两组间差异具有统计学意义(P<0.05),随着肺mPAP的增加,Th17/CD4+T细胞比例明显升高;Treg/CD4+T细胞的比例明显下降;Th17/Treg细胞比值显著升高。mPAP与Th17/Treg比值呈显著正相关(r=0.95,P<0.05)。结论:先天性心脏病合并肺动脉高压患者外周血中存在Th17/Treg失衡,且与肺动脉高压程度呈正相关,Th17/Treg失衡可能参与了肺动脉高压的发生发展。     

Treg和Th17细胞在寄生虫感染和卫生假说中的免疫调节

寄生虫感染在发展中国家仍然是一个严重威胁人类健康、影响社会经济发展的公共卫生问题。近年来,随着分子生物学和免疫学等研究进展,人们对寄生虫感染的免疫防御反应及免疫病理机制有了更多认识。CD4~+T细胞在机体免疫防御及免疫调节中发挥着非常重要的作用。传统上认为CD4~+T细胞在体内外可分化成Th1、Th2两大细胞亚群,它们产生不同的细胞因子,发挥不同功能。后来又发现了两种新的CD4~+T细胞亚群,即Th17细胞和调节性T(Treg)细胞。它们具有与传统的Th1、Th2细胞完全不同的、独立的分化和调节机制。已有大量研究证实,Treg和Th17细胞在寄生虫病的抗虫免疫及免疫病理机制中发挥着重要作用。此外,Th17和Treg细胞在寄生虫感染所诱导的卫生假说中的作用越来越受到重视。本文根据当前国内外有关Treg和Th17细胞分化及功能的研究进展,对Treg和Th17细胞在寄生虫感染及卫生假说中的作用作一简要综述。