隐球菌性脑膜炎患者外周血中Th9和Th17细胞水平变化及意义

目的：比较初发隐球菌性脑膜炎患者治疗前后与健康对照人群外周血 CD4＋ T 细胞中 Th9和 Th17细胞的比值,探讨 Th9和 Th17细胞在隐球菌性脑膜炎发病机制中的作用。方法选取初发未经治疗隐球菌性脑膜炎患者及健康对照各12例,抽取隐球菌性脑膜炎患者治疗前和治疗后3周及健康对照的外周血,分离外周血单核细胞,应用流式细胞仪检测技术对3组病例外周血 CD4＋ T 细胞中 Th9和 Th17的比值进行比较。结果与健康对照相比,隐球菌性脑膜炎患者治疗前Th17表达下调,差异有统计学意义;在治疗好转患者中,治疗后 Th17表达显著上调,与治疗前及健康对照相比差异均有统计学意义。Th9在治疗前与健康对照相比无差异,在治疗后隐球菌性脑膜炎患者中表达上调。结论 Th17免疫途径是隐球菌性脑膜炎患者抵御隐球菌感染的重要免疫机制,隐球菌性脑膜炎发病及治疗拮抗可能与 Th17缺乏有关。     

Th1、Th2和Th17型细胞在类风湿性关节炎和系统性红斑狼疮中的活化特点

探讨Th1、Th2和Th17型细胞在类风湿性关节炎(RA)和系统性红斑狼疮(SLE)发病机制中的作用。收集37例RA患者、25例SLE患者和34例健康人的抗凝血,应用ELISA检测血清中IFN-γ、IL-10和IL-17的水平。与健康对照组比较,RA和SLE患者血清中IFN-γ的水平均具有统计学意义(P<0.05);SLE患者IL-10水平出现有意义的升高(P<0.05);而RA患者IL-17的升高具有统计学意义(P<0.05)。由此提示Th1、Th2和Th17细胞在自身免疫性疾病中均发挥不同的重要作用。     

TLR2 agonist ameliorates murine experimental allergic conjunctivitis by inducing CD4 positive T-cell apoptosis rather than by affecting the Th1/Th2 balance

Innate immune responses that operate through Toll-like receptors (TLRs) are actively involved in the development of diseases predominantly mediated by adaptive immune responses. This is true also for allergic disease, as TLRs have been found to be involved in the development of allergic airway inflammation. We investigated whether stimulating TLR2 also abrogates murine allergic conjunctivitis by upregulating Th1 responses. We found that treating mice during the efferent phase with the TLR2 agonist Pam3CSK4 significantly suppressed eosinophil infiltration into the conjunctiva. However, Pam3CSK4 treatment inhibited both the Th1 and Th2 responses in the mice, and also suppressed eosinophil infiltration in IFN-gamma knockout mice. Flow cytometric analysis demonstrated that Pam3CSK4 treatment significantly elevated the numbers of annexin V-positive splenocytes, especially CD4 positive T cells. Thus, the stimulation of TLR2 during the efferent phase of murine allergic conjunctivitis suppresses eosinophil infiltration by inducing CD4 positive T-cell apoptosis rather than upregulating Th1 responses.     

Th22细胞与支气管哮喘

支气管哮喘是一种临床上常见的呼吸道疾病,研究发现CD4＋T细胞在哮喘的发病过程中起重要作用。Th22细胞是最近发现的一类CD4＋T细胞功能亚群之一,其主要效应因子IL-22在炎症性疾病、组织修复、创口愈合及自身免疫性疾病中起重要作用,但其具体机制尚未完全清楚。从Th22的细胞因子来源、生物学特性、分化和调控出发,简要探讨Th22细胞与哮喘之间的可能关系。     

异源反应性NK细胞对骨髓移植小鼠脾脏Th1/Th2细胞亚群的影响

目的:观察异源反应性自然杀伤细胞(NK细胞)对骨髓移植小鼠脾脏辅助性T细胞Th1/Th2亚群的影响。方法:以BALB/c和CB6F1小鼠为受体,在γ射线照射后通过尾静脉输入C57BL/6J小鼠的骨髓细胞和脾脏单个核细胞,建立移植物抗宿主病模型;然后输入供体的NK细胞,检测受体小鼠脾脏中Th1/Th2淋巴细胞亚群和外周血中γ干扰素(IFN-γ)、白细胞介素10(IL-10)的变化。结果:与单纯输入骨髓细胞和单个核细胞组小鼠相比,输入异源反应性NK细胞后,BALB/c和CB6F1小鼠脾脏中Th1细胞比例均下降,Th2细胞比例均上升,CB6F1小鼠脾脏中Th2虽有升高但没有统计学意义;外周血中IL-10水平显著升高,IFN-γ的水平显著下降。结论:异源反应性NK细胞可能通过降低脾脏中Th1细胞亚群比例和升高Th2细胞亚群比例减轻移植物抗宿主病。     

Leishmania donovani Infection of a Susceptible Host Results in Apoptosis of Th1-Like Cells: Rescue of Anti-Leishmanial CMI by Providing Th1-Specific Bystander Costimulation

A protective immune response against Leishmania donovani infection is mediated by T-helper type 1 (Th1) cells. Th1 induced cell-mediated immunity (CMI), as assessed by anti-leishmanial DTH response, is lost in a susceptible host such as BALB/c mice. Although the impaired Th1 function eventuates in unhindered parasite growth and in manifestation of the susceptible phenotype, the mechanism of down-regulation of the Th1 function is yet to be elucidated. Here, we provide evidence that the parasite downregulates the expression of a Th1-specific costimulatory molecule, M150, on the surface of infected BALB/c mice-derived macrophages. Th cells are rendered unresponsive to anti-CD3 Ab-mediated stimulation after interaction with infected macrophages. The anergized T cells produce much less IL-2, IL-4 and IFN-γ compared to those T cells which were costimulated using normal macrophages. The defect in proliferation, anti-CD3 Ab induced unresponsiveness and IFN-γ but not IL-4 production can be restored by providing bystander costimulation through M150. These results not only unfold a novel immune evasion strategy used by the parasite but also clarify the mechanism of Th1 cell debilitation during the disease. Recovery of Th1 cytokine production by bystander costimulation through M150 may help in formulating a new strategy for the elimination of intracellular parasites.     

Role of IL-27-producing dendritic [corrected] cells in Th1-immunity polarization in Lewis rats

Lewis and Brown Norway rats are entirely different with respect to the polarization of their immune responses (Th1 and Th2, respectively). We found that naive Lewis rat splenocytes treated in vitro with heat-killed Mycobacterium tuberculosis (Mtb) upregulate the expression of both subunits of IL-27 (IL-27p28 and EBI3). Mtb treatment caused naive Lewis rat splenocytes to express 4.6-fold more IL-27p28 than Mtb-treated Brown Norway rat splenocytes 6h after the treatment. Although WSX-1, the IL-27 receptor, was not induced by Mtb treatment in splenocytes from either rat strain, Lewis rats expressed significantly higher levels of the IL-27 signal transducers T-bet and IL-12Rbeta2 than Brown Norway rats. Flow cytometric analysis of dendritic cells from bone marrow cells revealed Lewis rats had more IL-27p28-positive cells. Thus, early in the immune response, Lewis rats appear to produce higher levels of IL-27 than Brown Norway rats, resulting in polarization towards Th1-immunity.     

分泌性中耳炎患者外周血Th1Th2细胞及T淋巴细胞亚群水平的表达及临床意义

目的:探讨分泌性中耳炎(SOM)患者外周血T辅助细胞1(Th1)、T辅助细胞2(Th2)细胞因子及T淋巴细胞亚群水平的表达及其临床意义。方法:选取我院于2015年1月至2018年1月期间收治的SOM患者135例记为SOM组,根据病程将患者分为急性组(病程14d,49例)、亚急性组(病程14-30d,53例)、慢性组(病程30d,33例)。另外选择同期于我院进行体检的100例健康者为对照组。分别对比SOM组和对照组受试者、不同病程SOM患者外周血Th1细胞因子[干扰素(INF-γ)、白细胞介素-2(IL-2)]、Th2细胞因子[白细胞介素-4(IL-4)、白细胞介素-10(IL-10)]以及T淋巴细胞亚群[CD3~+、CD4~+、CD8~+、CD4~+/CD8~+]水平。采用Pearson相关性分析INF-γ、IL-2、IL-4、IL-10与CD3~+、CD4~+、CD8~+、CD4~+/CD8~+的相关性。结果:SOM组患者外周血INF-γ、IL-2、IL-4、IL-10水平高于对照组(P0.05);急性组患者外周血INF-γ、IL-2、IL-4、IL-10水平低于亚急性组、慢性组,亚急性组患者外周血INF-γ、IL-2、IL-4、IL-10水平低于慢性组(P0.05)。SOM组患者外周血CD3~+、CD4~+、CD4~+/CD8~+水平低于对照组,CD8~+水平高于对照组(P0.05);急性组患者外周血CD3~+、CD4~+、CD4~+/CD8~+水平高于亚急性组、慢性组,CD8~+水平低于亚急性组、慢性组,亚急性组患者外周血CD3~+、CD4~+、CD4~+/CD8~+水平高于慢性组,CD8~+水平低于慢性组(P0.05)。Pearson相关性分析结果显示,SOM患者外周血INF-γ、IL-4与CD8~+呈正相关(P0.05),IL-4与CD3~+、CD4~+呈负相关(P0.05)。结论:SOM患者外周血Th1Th2细胞因子、T淋巴细胞亚群水平均表现异常,且其水平与疾病发生和发展存在一定联系,通过监测Th1Th2细胞因子、T淋巴细胞亚群有助于评估SOM患者病情。     

Th17细胞分化、调节及效应研究进展

Th17细胞作为一个不同于Th1、Th2的细胞亚群,已经被证实在自身免疫病、感染等疾病中发挥重要的作用.为了进一步认识Th17细胞的效应机制,近来对于Th17细胞的分化及调节进行了深入的研究,证实TGF-β与IL-6或者IL-21的协同作用是诱导Th17细胞分化的关键因素,而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.与Th1、Th2、Treg细胞特异性的转录调节因子T-bet、GATA3、Foxp3相对应,现证实ROR-γt(retinoid-related orphanreceptors-γr)是促进Th17细胞分化、调节其功能的特异性转录调节因子.Th17细胞通过分泌IL-17A、IL-17F、IL-21、IL-22、IL-6、TNF-α等细胞因子发挥效应功能.其中IL-21作为Th17细胞的一个自分泌调节凶子,在诱导Th17分化、抑制Th1、Treg功能方面发挥关键作用.而另一方面,近来发现,重要的T细胞生长因子IL-2在维持、促进Th1、Th2、Treg及CD8 T细胞功能活性的同时,却发挥着抑制Th17细胞分化的作用.Th1、Treg、Th17细胞的分化之间存在微妙的调节关系,TGF-β的水平、作用的时间决定着上述三群T细胞的分化结局.Th17细胞与Th1细胞均是自身免疫病及感染性疾病的重要效应细胞,二者的作用是否有时间、空间、功能方面的特异性?TGF-β如何调节两群效应细胞的分化方向及功能?以及Th17细胞在体内免疫平衡中的作用,是否可以通过Th17细胞诱导免疫耐受等,是人们急于回答的非常有意义的课题.     

创伤性骨折血清ALP、OPG表达与Th1/Th2平衡的相关性分析

摘要 目的：探讨创伤性骨折血清碱性磷酸酶（ALP）、骨保护素（OPG）表达与辅助性T细胞1（Th1）/辅助性T细胞2（Th2）平衡的相关性。方法：选取我院2019年2月到2022年12月收治的80例创伤性骨折患者作为研究对象,将其分为观察组,另选取同期来我院体检的80名健康志愿者作为对照组。抽取观察组患者入院后第二天清晨空腹静脉血和对照组清晨空腹静脉血,检测ALP、OPG、Th1、Th2表达水平,并分析血清ALP、OPG表达与Th1/Th2平衡的相关性。术后12个月后对观察组80例患者进行随访,依照患者骨折愈合程度将其分为骨折愈合组（n=54）和延迟愈合组（n=26）,对比两组患者临床一般情况与ALP、OPG、Th1/Th2表达水平,并分析创伤性骨折患者骨折愈合情况影响的多因素。结果：观察组与对照组ALP、OPG表达与辅助性T细胞水平对比差异显著,观察组ALP、OPG、Th2水平明显高于对照组,Th1、Th1/Th2水平明显低于对照组（P＜0.05）;Spearman相关分析结果表明：血清ALP、OPG与Th1/Th2平衡呈负相关（P<0.05）;骨折愈合组和延迟愈合组患者性别、年龄、致伤原因、Th2对比无明显差异（P＞0.05）,两组患者BMI、合并感染ALP、OPG、Th1、Th1/Th2水平对比差异显著（P＜0.05）;对上述单因素分析具有统计学差异指标进行赋值,多因素分析结果表明：ALP、OPG、Th2、Th1/Th2为影响创伤性骨折患者骨折愈合情况的独立影响因素（P＜0.05）。结论：创伤性骨折的发生会导致ALP、OPG水平升高,Th1/Th2平衡改变,患者血清ALP、OPG水平与Th1/Th2平衡关系具有明显相关性。另外,ALP、OPG、Th1、Th1/Th2水平变化可影响创伤性骨折患者术后骨折愈合水平。因此,临床上对于ALP、OPG水平升高,且Th1/Th2失衡患者需警惕骨折愈合不良的发生。     

Th17/Treg 细胞在银屑病的发病机制研究进展

Th17细胞和Treg细胞是CD4+T细胞在不同细胞因子环境中分化出的新亚群,发挥不同的生物学效应,使机体的免疫系统处于平衡状态.Th17/Treg细胞失衡可引起一系列自身免疫性疾病.银屑病是与遗传、免疫异常有关的皮肤炎症性疾病,其发病机制尚不清楚.越来越多的研究发现,Th17细胞增多和Treg细胞减少及其分泌的细胞因子在银屑病的发病中有着重要作用.本文围绕这一机制综述了近年来有关Th17细胞、Treg细胞在银屑病发病机制中作用的研究,帮助我们更深入地了解银屑病的发病机制并为今后临床诊断和治疗提供依据.     

慢性移植性肾病大鼠脾脏中辅助性T细胞和B 细胞免疫状态的研究*

目的:检测慢性移植性肾病(CAN)大鼠脾脏中辅助性T细胞(Th)和B细胞特征性因子表达量的变化,探究这些Th/B细胞免疫状态在CAN病程中的作用。方法:采用Fischer-Lewis左肾原位移植法建立大鼠慢性移植性肾病模型,Lewis-Lewis同种自体移植作为对照组。所有受体大鼠,术后8周处死,取脾脏组织,进行HE染色,拍照后采用双盲法评价脾脏组织病理变化程度及淋巴细胞浸润情况。用Trizol法提取脾脏组织中总RNA,采用实时荧光定量聚合酶链式反应(q RT-PCR)法检测各组脾脏中Th1,Th2,Th17,Treg和B细胞标志性因子的表达情况。结果:与对照组相比,CAN大鼠脾脏出现明显的结构肿胀及淋巴细胞浸润增多,并且Th1细胞特征性因子IFN-γ和T-bet表达量显著增加(P0.001,P0.05);Th2细胞特征性因子GATA-3表达升高(P0.001),但IL-4无变化;IFN-γ/IL-4比例明显上调(P0.001),T-bet/GATA3比例没有显著差异。Th17的特征性因子IL-17未见明显改变,而Treg细胞特征性因子Foxp3表达增加(P0.001),IL-17/Foxp3平衡明显向Treg细胞偏移(P0.05)。B细胞激活相关因子TNFRSF13C和RAG1表达量均显著上调(P0.01,P0.05),而RAG2水平则没有变化。结论:CAN大鼠脾脏中Th1/Th2的活性平衡向Th1偏移,分化平衡未出现显著变化;Th17/Treg的平衡向Treg细胞偏移,B细胞免疫状态也被激活,这些变化在CAN病程的发展中起到了重要作用,并且为临床监测和治疗提供了新的依据。     

沙门菌- 斑马鱼感染模型用于Th1/Th2 免疫应答的研究

目的:通过构建斑马鱼成鱼感染模型,研究鼠伤寒沙门菌感染对机体Th1/Th2免疫应答的影响。方法:用不同剂量细菌口饲感染8月龄的斑马鱼成鱼,绘制3周生存率曲线。观察各剂量下对成鱼的感染情况,并用实时荧光定量聚合酶链式反应检测肝脏Th1、Th2型细胞相关基因和细胞因子的mRNA转录水平,计算Th2/Th1相对表达量比值。结果:用105 CFU感染2周斑马鱼全部存活,第15天开始出现死亡,且在3周后死亡率达到50%;感染后3周解剖发现,肝脏、脾脏和肠道有明显红肿和糜烂;肝脏Th1、Th2型细胞相关基因和细胞因子mRNA转录水平明显向Th2偏移。结论:用105 CFU鼠伤寒沙门菌口饲感染斑马鱼成鱼构建的模型,能反映机体感染和免疫功能变化,可用于研究体内Th1/Th2免疫应答,为进一步研究鼠伤寒沙门菌感染与免疫机制提供了很好的实验工具。     

Murine NKT cells produce Th17 cytokine interleukin-22

Natural killer T (NKT) cells are known to produce Th17 cytokine IL-17 in addition to Th1/2 cytokines. In this study, the ability of NKT cells to produce IL-22, another Th17 cytokine, was examined in mice. When murine spleen cells were stimulated with α-galactosyl ceramide, a ligand for NKT cells, not only Th1/2 cytokines (IFN-γ, IL-4) but Th17 cytokines (IL-17, IL-22) were produced. NKT cells isolated from splenocytes released IL-17 and IL-22 following CD3, CD3/IL-2 or CD3/CD28 stimulation, in which CD3/CD28 costimulation was most effective. Production of IL-17 and IL-22 in CD4+ and CD8+ T cells from splenocytes was little, if any, even after CD3/CD28 costimulation. Treatment with IL-6/TGF-β decreased CD3/CD28-stimulated production of IL-22, but not that of IL-17, in NKT cells. These findings show for the first time that NKT cells are a cell source of IL-22, and that expression of two Th17 cytokines might be regulated in NKT cells by different mechanisms.     

In vitro Th1 cytokine-independent Th2 suppressive effects of bifidobacteria

A comparison between 17 strains of lactic acid bacteria and 15 strains of bifidobacteria indicated that bifidobacteria induced significantly lower levels of interleukin-12 (IL-12) in murine splenic cells. The present study aims to evaluate the effect and mechanism of Bifidobacterium longum BB536, a probiotic strain, in suppressing antigen-induced Th2 immune response in vitro. BB536 suppressed immunoglobulin (Ig) E and IL-4 production by ovalbumin-sensitized splenic cells, but induction of Th1-inducing cytokine production, such as IL-12 and gamma interferon (IFN-gamma) tended to be lower compared with lactic acid bacteria. Neutralization with antibodies to IL-12, IFN-gamma, IL-10 and transforming growth factor beta indicated negative involvement of Th1-inducing cytokines and regulatory cytokines in the suppression of Th2 immune response by BB536, especially when treated at higher doses of BB536 (>10 microg cells/ml). Furthermore, BB536 induced the maturation of immature bone marrow-derived dendritic cells (BM-DCs), and suppressed antigen-induced IL-4 production mediated by BM-DCs. These results suggested that BB536 suppressed Th2 immune responses, partially independent of Th1-inducing cytokines and independent of regulatory cytokines, mediated by antigen-presenting cells such as dendritic cells.     

Exosome derived from CD137‐modified endothelial cells regulates the Th17 responses in atherosclerosis

The role of exosomes derived from endothelial cells (ECs) in the progression of atherosclerosis (AS) and inflammation remains largely unexplored. We aimed to investigate whether exosome derived from CD137‐modified ECs (CD137‐Exo) played a major role in AS and to elucidate the potential mechanism underlying the inflammatory effect. Exosomes derived from mouse brain microvascular ECs treated with agonist anti‐CD137 antibody were used to explore the effect of CD137 signalling in AS and inflammation in vitro and vivo. CD137‐Exo efficiently induced the progression of AS in ApoE^?/? mice. CD137‐Exo increased the proportion of Th17 cells both in vitro and vivo. The IL‐6 contained in CD137‐Exo which is regulated by Akt and NF‐КB pathway was verified to activate Th17 cell differentiation. IL‐17 increased apoptosis, inhibited cell viability and improved lactate dehydrogenase (LDH) release in ECs subjected to inflammation induced by lipopolysaccharide (LPS). The expression of soluble intercellular adhesion molecule1 (sICAM‐1), monocyte chemoattractant protein‐1 (MCP‐1) and E‐selectin in the supernatants of ECs after IL‐17 treatment was dramatically increased. CD137‐Exo promoted the progression of AS and Th17 cell differentiation via NF‐КB pathway mediated IL‐6 expression. This finding provided a potential method to prevent local and peripheral inflammation in AS.     

Ursolic acid inhibits Th17 cell differentiation via STAT3/RORγt pathway and suppresses Schwann cell-mediated Th17 cell migration by reducing CXCL9/10 expression

Th17 cells represent important immune cells. Ursolic acid (UA) can regulate immune cell activities. This study was aimed to explore the effects of UA on Th17 cell differentiation and Schwann cell(SCs)-mediated migration and the potential mechanism. Naïve CD4⁺ T cells were isolated from rat peripheral blood, induced for Th17 cell differentiation, and treated with UA. The proportion of Th17 cells was detected by flow cytometry assay. SCs were co-cultured with Th17 cells. Th17 cell migration was detected by Transwell assay. The molecule expression was determined by Western blot and qRT-PCR. UA inhibited the Th17 cell differentiation and suppressed the STAT3/RORγt pathway. STAT3 overexpression up-regulated p-STAT3 and RORγt expression and promoted Th17 cell differentiation under the UA treatment. In LPS- and IFN-γ-stimulated-SCs, UA suppressed the expression of chemokines CXCL9/10, but had no significant effect of CCL20 expression. The expression CXCL9/10 receptor CXCR3 was higher in Th17 cells than that in Treg cells, while the expression CCL20 receptor CCR6 was lower in Th17 cells than that in Treg cells. UA, anti-CXCR3, and anti-CCR6 treatment inhibited SCs-mediated Th17 cell migration, and anti-CXCR3 exerted stronger inhibitory effect than Anti-CCR6. UA inhibited Th17 cell differentiation through STAT3/RORγt pathway and suppressed Th17 cell migration through down-regulating CXCL9/10 expression in SCs.     

Th17 immune response to adipose tissue-derived mesenchymal stromal cells

Adipose tissue-derived mesenchymal stromal cells (ASCs) hold the promise of achieving successful immunotherapeutic results due to their ability to regulate different T-cell fate. ASCs also show significant adaptability to environmental stresses by modulating their immunologic profile. Cell-based therapy for inflammatory diseases requires a detailed understanding of the molecular relation between ASCs and Th17 lymphocytes taking into account the influence of inflammation and cell ratio on such interaction. Accordingly, a dose-dependent increase in Th17 generation was only observed in high MSC:T-cell ratio with no significant impact of inflammatory priming. IL-23 receptor (IL-23R) expression by T cells was not modulated by ASCs when compared to levels in activated T cells, while ROR-γt expression was significantly increased reaching a maximum in high (1:5) unprimed ASC:T-cell ratio. Finally, multiplex immunoassay showed substantial changes in the secretory profile of 15 cytokines involved in the Th17 immune response (IL-1β, IL-4, IL-6, IL-10, IL-17A, IL-17F, IL-22, IL-21, IL-23, IL-25, IL-31, IL-33, IFN-γ, sCD40, and TNF-α), which was modulated by both cell ratio and inflammatory priming. These findings suggest that Th17 lymphocyte pathway is significantly modulated by ASCs that may lead to immunological changes. Therefore, future ASC-based immunotherapy should take into account the complex and detailed molecular interactions that depend on several factors including inflammatory priming and cell ratio.