IL-2对免疫激活和免疫耐受的双向调节作用

白细胞介素2(interleukin-2,IL-2)从被发现迄今已有30余年,但仍然是最受关注和被广泛研究的细胞因子之一。IL-2最初是从T细胞培养上清液中分离获得,并对体外培养的T细胞具有促进增殖作用,因此传统认为IL-2是一种激活T细胞,并维持T细胞分化和增殖的T细胞生长因子,同时也参与炎症或自身免疫性反应。近年来,人们在IL-2或IL-2R缺陷小鼠动物模型等的研究中发现IL-2的主要功能不仅是提升免疫反应,更重要的是维持Treg细胞的稳定及其介导的免疫耐受,故具有双向免疫调节作用,这些对IL-2生物学活性的新发现,也促使人们开始重新评价和认识临床使用IL-2的疗效、适应症和给药方式。     

供体特异性调节性T细胞在肝移植免疫耐受中的意义

免疫耐受是器官移植的终极目标。尽管目前对于移植术后免疫耐受的探索很多,但大多处于试验阶段,应用于临床的方案甚少。对调节性 T细胞诱导免疫耐受的研究较为透彻,但供体特异性调节性 T细胞因其比例较低、制备困难,在移植免疫中的研究相对较少。本文从调节性 T细胞的共性、供体特异性调节性 T细胞的抑制功能、体外培养技术、胸腺移植等方面出发,总结供体特异性调节性 T 细胞在近年的研究热点,以期为临床研究提供新思路。     

老年原发免疫性血小板减少症患者治疗前后T淋巴细胞亚群的临床研究

目的 检测老年原发免疫性血小板减少症(ITP)患者治疗前后T淋巴细胞亚群的动态变化, 探讨其在ITP发生发展中的作用。方法 采用流式细胞术测定ITP患者治疗前后及正常对照组外周血T淋巴细胞亚群的水平。结果 ITP患者治疗前后T淋巴细胞绝对值、CD3⁺、CD4⁺、CD8⁺、CD4⁺CD25⁺T淋巴细胞比例及CD4⁺/CD8⁺比值分别为(0.83±0.16)vs(1.74±0.36)、(71.71±1.07)% vs(72.69±1.35)%、(41.78±0.71)% vs(42.46±1.20)%、(29.67±0.97)% vs(28.56±1.75)%、(8.76±0.56)% vs(9.39±1.26)%、(1.42±0.07)vs(1.49±0.13), CD8⁺T淋巴细胞比例治疗后显著降低, 其余均显著升高, 差异有统计学意义(P<0.05)。结论 T淋巴细胞亚群的异常改变, 破坏自身免疫, 与病情相关, 可指导临床治疗, 并作为评估预后的参考指标。     

Th17／Treg 失衡在肝纤维化中的研究进展

肝纤维化是一种机体的慢性炎症反应,其特征是细胞外基质（ECM）合成与降解的失衡,从而造成肝内 ECM的病理性沉积,导致肝功能紊乱,最终发展为肝硬化,甚至是肝癌。肝纤维化的发病机理目前尚未清楚。随着免疫学的发展,T 细胞在肝纤维化的发病过程中的作用逐步得到认识,Th17细胞与 Treg 细胞这两种 T 细胞亚型在肝纤维化的发病过程中的作用已逐渐被认可。因此,该文就这两种细胞以及这两种细胞的相互作用在肝纤维化过程中的研究进展进行综述。     

LncRNA GAS5 inhibits Th17 differentiation and alleviates immune thrombocytopenia via promoting the ubiquitination of STAT3

BackgroundThe increased differentiation of T helper 17 cells (Th17) accelerates the development of immune thrombocytopenia (ITP), which is a common autoimmune disease with limited therapeutic methods. Recent studies have revealed that long non-coding RNAs (lncRNAs) play a critical role in autoimmune diseases, thus this study aims to investigate the effect of lncRNA GAS5 on the differentiation of Th17 cells in ITP.MethodsThe expression of GAS5 in peripheral blood mononuclear cells (PBMCs) of ITP patients and spleen tissues of ITP mice was measured by qRT-PCR. The percentage of Th17 cells in CD4⁺ cells was measured by flow cytometry. The combination between GAS5 and STAT3 was confirmed by RNA pull-down assay and RNA Binding Protein Immunoprecipitation (RIP). The ubiquitination of STAT3 was detected by ubiquitination assay and the interaction between STAT3 and TRAF6 was measured by Co-Immunoprecipitation (Co-IP). Finally, the effect of GAS5 on Th17 differentiation was investigated in vitro and in vivo using lentivirus (lenti)-GAS5.ResultsGAS5 expression was downregulated both in PBMCs of ITP patients and spleen tissues of ITP mice. Overexpression of GAS5 suppressed Th17 differentiation while had no effect on Treg differentiation in naïve CD4⁺ cells. RNA pull-down and RNA immunoprecipitation assays confirmed the interaction between GAS5 and STAT3. Further studies showed GAS5 accelerated the degradation of STAT3 via promoting TRAF6-mediated ubiquitination. Overexpressing GAS5 suppressed Th17 differentiation in vitro and alleviated ITP in vivo via reducing STAT3.ConclusionLncRNA GAS5 inhibited Th17 differentiation through promoting the TRAF6-mediated ubiquitination of STAT3, thus relieving ITP.     

调节性B细胞在炎症免疫相关疾病的调控作用及机制

调节性免疫细胞具有广义和狭义概念,狭义上,调节性免疫细胞是指具有负性调节作用的免疫细胞。调节性B细胞(regulatory B cell,Breg)在控制炎症免疫相关疾病免疫应答、介导免疫耐受中可能发挥重要作用。Breg通过分泌抑制性细胞因子IL-10和TGF-β负性调控炎症免疫相关疾病的病理过程。TLRs/MyD88信号转导通路在介导Breg功能中起到关键作用。TLRs/MyD88信号转导活化促进Breg细胞产生IL-10,抑制T细胞活化及IFN-γ分泌。不同TLR控制着炎症免疫相关疾病的启动和恢复。运用优先触发Breg细胞调节功能的TLR激动剂,分离TLRs的相反作用,对控制免疫反应,制定合理的炎症免疫相关疾病治疗策略具有重要的意义。     

Downregulation of regulatory T cell function in patients with delayed fracture healing

Bone fracture healing is a multistage regenerative process that requires the collaboration of various cell types, with approximately 5%‐10% of fractures not healing properly. Accumulating evidence suggests that dysregulations in the immune system are associated with defective healing. In a cohort of 30 bone fracture patients between 50 and 62 years of age, 8 patients displayed delayed healing. Compared to the 22 normal healing patients, these 8 delayed healing patients presented significantly lower frequencies of CD4⁺CD25^hiFoxp3⁺ canonical regulatory T cells immediately following bone fracture and early on during the healing process. The CD4⁺CD25^+/hi T cells from delayed healing patients also presented reduced capacity to express transforming growth factor beta (TGF‐β), and presented reduced surface expression levels of inhibitory molecules, including CTLA‐4 and Lag‐3, compared to CD4⁺CD25^+/hi T cells from normal healing patients. Moreover, CD4⁺CD25^+/hi T cells from delayed healing patients were less potent in the suppression of CD4⁺CD25^? autologous conventional T cell proliferation, and presented reduced expansion capacity in response to interleukin (IL)‐2 stimulation. Overall, our results demonstrated multiple reductions in regulatory T cell function in delayed healing patients that could produce long‐lasting consequences in the bone fracture healing process.     

凉血解毒方药对原发免疫性血小板减少症患者外周血CD4＋CD2＋5调节性 T 细胞水平的影响

目的：观察原发免疫性血小板减少症（ ITP）患者治疗前后外周血CD4＋CD2＋5调节性T细胞水平变化,探讨凉血解毒方药对ITP患者CD4＋CD2＋5调节性T细胞的作用。方法选取56例ITP患者,男12例,女44例;年龄18～62岁,中位年龄29岁。给以凉血解毒方药治疗（地黄止血胶囊2‘．0 g,3次／d,口服,凉血解毒中草药煎剂升板汤,1剂／d）。患者于治疗前、治疗后90 d分别采取外周静脉血,采用流式细胞术检测CD4＋CD2＋5调节性T细胞水平。20例健康体检者为正常对照组。结果治疗后第90天,总有效率为73．2％。56例ITP患者治疗前外周血 CD4＋CD2＋5调节性T细胞表达水平低于正常对照组[（1．01±0．67）％,（2．81±0．52）％],差异有统计学意义（ P ＜0．05）;重症ITP患者CD4＋CD2＋5调节性 T细胞水平低于非重症患者[（0．71±0．23）％,（1．48±0．64）％],差异有统计学意义（ P ＜0．01）;患者治疗后90 d CD＋4 CD2＋5调节性T细胞水平高于治疗前[（1．93±0．53）％,（10．1±0．67）％],与治疗前比较差异有统计学意义（ P ＜0．05）;治疗后有效组CD4＋CD2＋5调节性T细胞水平高于无效组,差异有统计学意义（P ＜0．05）。结论 ITP 患者外周血CD 4＋CD 2＋5调节性 T 细胞水平低于正常对照组（ P ＜0．05）;凉血解毒方药提升CD4＋CD2＋5调节性T 细胞水平可能是治疗ITP的疗效机制。     

Ursolic acid ameliorates autoimmune arthritis via suppression of Th17 and B cell differentiation

Aim:

Ursolic acid (UA) is a pentacyclic triterpenoid found in most plant species, which has been shown anti-inflammatory and anti-oxidative activities. In this study, we examined the effects of UA on collagen-induced arthritis (CIA) in mice, and to identify the mechanisms underlying the effects.

Methods:

CIA was induced in mice. Two weeks later, the mice were treated with UA (150 mg/kg, ip, 3 times per week) for 4 weeks. The expression of cytokines and oxidative stress markers in joint tissues was measured with immunohistochemistry. The numbers of CD4⁺IL-17⁺, CD4⁺CD25⁺Foxp3⁺ and pSTAT3 cells in spleens were determined using confocal immunostaining or flowcytometric analyses. Serum antibody levels and B cell-associated marker mRNAs were analyzed with ELISAs and qRT-PCR, respectively. CD4⁺ T cells and CD19⁺ B cells were purified from mice spleens for in vitro studies.

Results:

UA treatment significantly reduced the incidence and severity of CIA-induced arthritis, accompanied by decreased expression of proinflammatory cytokines (TNF-α, IL-1β, IL-6, IL-21 and IL-17) and oxidative stress markers (nitrotyrosine and iNOS) in arthritic joints. In CIA mice, UA treatment significantly decreased the number of Th17 cells, while increased the number of Treg cells in the spleens, which was consistent with decreased expression of pSTAT3, along with IL-17 and RORγt in the splenocytes. In addition, UA treatment significantly reduced the serum CII-specific IgG levels in CIA mice. The inhibitory effects of UA on Th17 cells were confirmed in an in vitro model of Th17 differentiation. Furthermore, UA dose-dependently suppressed the expression of B cell-associated markers Bcl-6, Blimp1 and AID mRNAs in purified CD19⁺ B cells pretreated with IL-21 or LPS in vitro.

Conclusion:

UA treatment significantly ameliorates CIA in mice via suppression of Th17 and differentiation. By targeting pathogenic Th17 cells and autoantibody production, UA may be useful for the treatment of autoimmune arthritis and other Th17-related diseases.     

sST2/IL-33与Th17/Treg相关细胞因子在原发免疫性血小板减少症中的作用研究

目的 观察可溶性ST2（sST2）和白细胞介素（IL）-33在原发免疫性血小板减少症（ITP）中的表达水平变化及其对Th17/Treg细胞平衡的调节机制。方法 30例ITP患者作为ITP组,选取同期健康体检者20例作为对照组,采用酶联免疫吸附试验（ELISA）检测 2组外周血血清中 IL-33、sST-2、IL-17和转化生长因子-β（TGF-β）的水平。结果 与对照组比较,ITP组血清sST2及IL-17水平均升高（P＜0.01）,TGF-β表达水平降低（P＜0.01）,而IL-33的表达水平差异无统计学意义（P＞0.05）;在 ITP 组中 sST2 与 TGF-β 呈负相关（r=-0.471,P＜0.01）,与 IL-17 无相关性（r=0.189,P＞0.05）。结论 sST2表达水平增加和Th17/Treg细胞失衡可能是导致ITP发病的免疫机制之一。     

泼尼松联合长春新碱治疗慢性免疫性血小板减少症的疗效及对细胞免疫功能的影响

目的　研究泼尼松联合长春新碱治疗慢性免疫性血小板减少症的疗效及对细胞免疫功能的影响。方法　选取中山市人民医院2019年1月至2021年8月期间收治的136例慢性免疫性血小板减少症患儿为研究对象,进行前瞻性研究,按照随机数字表法等分为两组,分别予以不同的药物进行治疗。对照组中男33例,女35例,年龄（11.13±2.83）岁,采取泼尼松治疗;研究组中男34例,女34例,年龄（11.12±2.28）岁,在泼尼松治疗的基础上联合长春新碱治疗。持续治疗2个月后比较两组患者治疗前后的细胞免疫功能变化情况以及不同疗法的治疗效果和不良反应发生概率。使用SPSS 22.0统计软件分析数据,采用t、χ²检验。结果　治疗后,研究组的CD3⁺、CD4⁺、CD4⁺/CD8⁺分别为（54.87±4.59）%、（33.97±3.85）%、（1.85±0.36）,均高于对照组［分别为（52.35±5.24）%、（31.03±3.70）%、（1.72±0.21）］,研究组CD8⁺为（23.78±2.38）%,低于对照组的（25.32±2.43）%,两组比较差异均有统计学意义（t=2.983、4.720、2.572、3.733,均P<0.05）。研究组患者的治疗有效率为94.12%（64/68）,高于对照组的79.41%（54/68）,两组比较差异有统计学意义（χ²=6.403,P=0.011）。两组不良反应发生情况差异无统计学意义（χ²=0.119,P=0.730）。结论　泼尼松联合长春新碱治疗慢性免疫性血小板减少症患儿具有较好的效果,并且对患儿的细胞免疫功能具有积极影响,值得广泛推广。     

IL-17 cytokine/receptor families: emerging targets for the modulation of inflammatory responses

IL-17 was identified a decade ago as a pro-inflammatory cytokine produced by activated T cells that stimulates the secretion of other cytokines from various non-lymphoid cells by acting through a unique cell surface receptor, IL17R. Evidence that IL-17 may contribute to several immune-mediated diseases, such as rheumatoid arthritis and airway inflammation, prompted much interest in this cytokine. Recently, the large-scale analysis of expressed sequence tags (EST) led to the discovery of novel genes dispersed in the human genome that encode at least five additional cytokines structurally related to IL-17. Screening of EST databases also uncovered at least four novel genes encoding Type I transmembrane proteins with significant homology to IL-17R, thereby forming a family of receptors whose cognate ligands are likely to belong to the IL-17 cytokine family. Initial characterisation of some of these cytokines and one IL-17R homologue demonstrated their involvement in regulating inflammatory responses in a manner similar to, albeit distinct from, that of prototypic IL-17. The IL-17 cytokine/receptor families appear therefore to represent unique signalling systems within the cytokine network that might offer innovative approaches to manipulate immune and inflammatory responses. The prospect of targeting these molecules for therapeutic purposes has generated a substantial volume of patent literature that will be reviewed here.     

氯雷他定对变应性鼻炎患儿外周血 T细胞17的影响

目的：观察氯雷他定对变应性鼻炎患儿外周血T细胞17（Th17）阳性率及白细胞介素（ IL）－17及IL－23水平的影响。方法试验组为80例急性发作期变应性鼻炎患儿,每日给予氯雷他定糖浆5 mL,睡前口服,2周为1个疗程;对照组为30例健康儿童,不予任何药物。用流式细胞术及酶联免疫吸附试验（ELISA）法检测试验组治疗前、治疗后1,2周以及对照组的外周血Th17细胞阳性率及IL－17、IL－23水平。结果试验组患儿治疗前血清中Th17细胞阳性率及IL－17、IL－23的含量较对照组明显升高（P＜0．05）。治疗2周后,Th17细胞阳性率及IL－17、IL－23的含量较对照组仍升高,但差异缩小（ P＜0．05）。治疗1周后,试验组Th17细胞阳性率及IL－17、IL－23的含量较治疗前降低,差异无统计学意义（ P ＞0．05）。治疗2周后,试验组 Th17细胞阳性率及 IL －17、IL－23的含量较治疗前明显降低（ P＜0．05）。结论氯雷他定糖浆可以通过调节患者外周血Treg／Th17平衡,降低IL－17和IL－23 mRNA的表达水平来抑制变应性鼻炎的炎症反应。     

CD4+IL-10+调节性T细胞在特应性皮炎患者中的改变

摘要 目的 探讨特应性皮炎患者CD4+IL-10+调节性T细胞及其细胞因子的改变。方法 用流氏细胞仪直接免疫荧光法检测AD患者外周血CD4+IL-10+T细胞的绝对计数和百分率;用酶联免疫吸附试验检测外周血细胞因子IL-10、TGF-β的浓度。 结果AD患者CD4+IL-10+T细胞绝对计数为0.26+0.126×109/L,占CD4+T细胞的百分率是12.76+4.85%;而对照组CD4+IL-10+T细胞绝对计数为0.016+0.007×109/L,占CD4+T细胞的百分率是3.27+1.03%。经t检验,AD患者CD4+IL-10+T细胞绝对计数和百分率均高于对照组,有统计学差异（p <0.05）。 AD患者外周血IL-10浓度为126.59+59.46 pg/ml,高于正常对照组（31.47+10.96 pg/ml）,经t检验,有统计学差异（p<0.05）。AD患者外周血TGF-β浓度为407.23+155.88 pg/ml,高于正常对照组（359.47+99.21 pg/ml）,但经t检验,没有统计学差异（p>0.05）。CD4+IL-10+T细胞绝对计数、百分率均与外周血IL-10浓度呈正相关,有统计学意义（r=0.47,r=0.65, p <0.05）;而与外周血TGF-β无统计学相关性（r=0.15,r=0.21 , p >0.05）。结论 IL-10可能是Trl发挥功能的主要因子,通过下调由Thl和Th2细胞介导的炎症反应过程,参与免疫调节。     

The JAK2 inhibitor AG490 regulates the Treg/Th17 balance and alleviates DSS-induced intestinal damage in IBD rats

The pathogenesis of inflammatory bowel disease (IBD) remains unclear, and it is currently believed that an imbalance in regulatory T (Treg) cells/T helper 17 cells (Th17 cells) is related to the occurrence and development of IBD. Recently, the JAK2 inhibitor AG490 has been used in animal models such as rheumatoid arthritis and bronchial asthma models and shown to exert immunoregulatory functions that improve disorder in the Treg/Th17 cell balance. This study aimed to evaluate the effect of AG490 on the intestinal inflammatory process in an IBD rat model. A dextran sulfate sodium (DSS)-induced IBD rat model was established, and disease activity index (DAI) scores were calculated. The histopathological damage score was determined by haematoxylin-eosin (H&E) staining. Treg/Th17 cells in the spleen were detected by flow cytometry. The levels of interleukin (IL)-10, IL-6 and IL-17A were detected by enzyme-linked immunosorbent assay (ELISA). AG490 attenuated DSS-induced IBD injury by regulating the Treg/Th17 balance and related cytokine secretion to reduce the DAI and colonic tissue damage. Thus, AG490 may be a new method for effective treatment of IBD.     

Indoles mitigate the development of experimental autoimmune encephalomyelitis by induction of reciprocal differentiation of regulatory T cells and Th17 cells

Background and Purpose

Dietary indole derivatives, indole-3-carbinol (I3C) and diindolylmethane (DIM), possess anti-cancer properties and exhibit the characteristics of aryl hydrocarbon receptor (AhR) ligands. Because AhR activation has recently been shown to regulate T cell differentiation, we tested the hypothesis that I3C and DIM may mediate anti-inflammatory properties by promoting regulatory T cell (T-regs) differentiation while inhibiting Th17 cells.

Experimental Approach

We investigated the therapeutic efficacy of I3C and DIM against experimental autoimmune encephalomyelitis (EAE), a murine model of multiple sclerosis (MS). The efficacy was evaluated based on clinical scores of paralysis, histopathology, serum cytokines and infiltration of T cells in the CNS. We next studied the mechanism of induction of T cells against myelin oligodendrocyte glycoprotein (MOG_35–55) peptide, both in vivo and in vitro, specifically investigating the differentiation of T-regs and Th17 cells, and determined if indoles were acting through AhR.

Key Results

Pretreatment of EAE mice with I3C or DIM completely prevented the clinical symptoms and cellular infiltration into the CNS. Also, post-treatment of EAE with I3C or DIM proved highly effective in curtailing the overall severity of the disease. In addition, I3C or DIM promoted the generation of T-regs, while down-regulating the induction of MOG-specific Th17 cells. The regulation of FoxP3 induction and suppression of Th17 cells by indoles in vivo and in vitro were found to be AhR-dependent.

Conclusions and Implications

Together, our studies demonstrate for the first time that I3C and DIM may serve as novel therapeutics to suppress neuroinflammation seen during MS through activation of AhR.     

中药调控调节性T细胞改善缺血性脑卒中后免疫功能的研究进展

在缺血性脑卒中病理进程中,免疫炎症反应贯穿全程,是导致神经元凋亡与坏死的重要原因,与T淋巴细胞亚群的紊乱关系密切。调节性T细胞是一类能发挥免疫调节作用的T细胞亚群,对缺血性脑卒中的发生发展有重要影响。中药具有多成分、多靶点、多通路的特征,可通过调控调节性T细胞数量,纠正辅助性T细胞17与调节性T细胞失衡状态,从而抑制炎症反应,对抗动脉粥样硬化,减轻神经功能缺损,提高患者免疫功能并改善预后,对缺血性脑卒中及其并发症的发生发展起到防治作用。本文综述了中药调控调节性T细胞改善缺血性脑卒中后免疫功能方面的研究,以期为中药治疗缺血性脑卒中的免疫学研究提供新思路。     

IL-23／IL-17轴对寻常型银屑病病情的影响

目的：通过观察甲砜霉素联合异维A酸胶丸治疗寻常型银屑病的疗效,检测患者血清辅助T细胞17（ Th17）相关因子白介素17（IL-17）、白介素22（IL-22）和白介素23（IL-23）水平的变化,分析其与疾病严重程度及转归的关系,探讨银屑病可能的发病机制。方法治疗组为寻常型银屑病40例,对照组为健康体检者40例;治疗组给予甲砜霉素、异维A酸胶丸口服及丁酸氢化可的松乳膏外用。以4周为1个疗程,2个疗程后进行疗效判定。治疗组治疗前后分别检测血清IL-17、IL-22和IL-23的水平,与对照组进行比较,分析IL-17、IL-22、IL-23与严重程度指数（psoriasis area and severity index,PASI）相关性。结果治疗组治疗前后血清IL-17、IL-22、IL-23水平较对照组高,差异有统计学意义（P＜0．01）,治疗8周后IL-17、IL-22、IL-23水平较治疗前明显下降（ P＜0.01）。治疗组治疗前IL-17、IL-22与IL-23表达水平与PASI评分之间均呈正相关（ r分别为0.77、0．76、0．60,P均＜0．05）。结论 IL-23／IL-17轴可能在寻常型银屑病的发病机制中起重要作用,IL-17、IL-22、IL-23可以作为判断寻常型银屑病严重程度的重要指标,降低IL-17、IL-22、IL-23表达可能成为治疗寻常型银屑病的新的方向。