Th17细胞及其相关因子在银屑病发病中的作用及其分子机制

银屑病是一种自身免疫性疾病,表现为以T细胞为主的免疫功能紊乱。Th17细胞是一种能分泌白细胞介素17(IL-17)的T细胞亚群,参与多种自身免疫性疾病的免疫调节。研究发现银屑病患者外周血中存在IL-23和Th17类细胞因子的显著升高,IL-23/Th17细胞轴在银屑病发病机制中有重要作用。除Th17细胞本身,在银屑病患者皮损处,IL-17、IL-22、IL-21等Th17相关细胞因子亦发生改变。调节性B细胞在银屑病的发病过程中有可能通过产生IL-10发挥负向免疫调节功能。本文主要从Th17细胞及其相关因子与银屑病,调节性B细胞和Th17细胞在银屑病发病机制中的拮抗作用几个方面进行综述。     

Graves病小鼠模型IL-10水平与Th17细胞功能的检测和意义

目的 研究Th17细胞及相关细胞因子IL-17 mRNA与IL-10 mRNA在实验性Graves小鼠体内变化.方法 利用TSHR-E(胞外区)的腺病毒免疫小鼠方法建立Graves疾病模型,化学发光法检测血清FT4水平,细胞培养生物学方法检测TSAb活性,流式细胞术(FCM)分析小鼠脾脏细胞中Th17细胞的比例,qRT-PCR检测小鼠脾脏细胞中IL-17和IL-10mRNA水平.结果 小鼠血清FT4及TSAb水平升高,TSAb与FT4水平呈显著正相关(r=0.90,P＜0.01);Graves小鼠脾细胞中Th17细胞的比例为(2.56±0.43)％,明显高于正常对照组小鼠(1.07±0.27％)％(P＜0.05);Graves小鼠脾细胞中IL-17 mRNA的含量明显高于正常对照组(P＜0.05),而IL-10 mRNA的含量明显低于正常对照组(P＜0.05);且IL-17和IL-10水平变化存在明显的负相关(r=-0.564,P＜0.01).结论 TSHR-E(胞外区)的腺病毒免疫建立的GD模型中,IL-17mRNA增加和IL-10 mRNA则明显下降;提示两者可能在GD的发生及发展中发挥免疫调节作用.     

IL-23/IL-17轴在银屑病免疫发病机制中的作用

银屑病是一种以T淋巴细胞异常活化和浸润为主要特征的慢性炎性反应皮肤病。Th17细胞及IL-23/IL-17轴在银屑病的发病机制中可能处于关键地位,并成为新的治疗靶标。IL-23诱导Th17细胞分化增殖,分化成熟的Th17可以分泌IL-17、IL-21、IL-22等多种细胞因子,Th17类细胞因子在银屑病等多种自身免疫疾病和炎性疾病中起重要作用。     

商陆皂苷甲降低银屑病模型小鼠Th17细胞相关细胞因子的表达

目的观察商陆皂苷甲(EsA)对银屑病小鼠17型辅助T(Th17)细胞相关细胞因子表达的影响。方法将48只雌性BALB/c小鼠随机分为空白对照组、模型组、退银汤组[66.60 g/(kg·d)]、 EsA低剂量组[5 mg/(kg·d)],EsA中剂量组[10 mg/(kg·d)],EsA高剂量组[20 mg/(kg·d)],每组8只。采用咪喹莫特乳膏诱导小鼠银屑病模型,以皮损面积和严重程度指数(PASI)评分为标准,观察银屑病样小鼠皮损的变化情况; HE染色观察皮损组织形态学变化,ELISA检测小鼠血清Th17细胞相关细胞因子白细胞介素17(IL-17)、IL-22、 IL-6、肿瘤坏死因子α(TNF-α)的水平。结果与模型组相比,退银汤组、 EsA各组PASI评分降低,退银汤组与EsA高剂量组降低较明显;各用药组对银屑病样小鼠背部皮肤组织病理变化减轻;与空白对照组相比,模型组小鼠Th17细胞相关细胞因子水平显著升高,各用药组细胞因子水平明显低于模型组,其中EsA高剂量组IL-6含量接近于空白对照组。结论 EsA可能通过降低Th17细胞相关细胞因子的表达改善银屑病样小鼠皮损状况。     

利用基因敲除小鼠研究咪喹莫特诱导银屑病模型中转录因子RORγt的作用

目的观察RORγt转录因子在咪喹莫特诱导的小鼠银屑病模型中的作用。方法雌性8周C57BL/6小鼠随机分为正常对照组、野生型给药组、RORγt-/-给药组,用PASI评分观察小鼠模型皮损的变化,HE染色观察皮损组织形态学变化,实时荧光定量PCR对小鼠皮损组织中的细胞因子mRNA相对表达量进行检测。结果局部给予咪喹莫特后野生型小鼠背部皮肤出现红斑、鳞屑、增厚典型银屑病样皮损,RORγt-/-给药组小鼠症状明显减轻。RORγt-/-组IL-22、IL-17、IL-23细胞因子的相对表达量明显低于野生型给药组。结论 RORγt转录因子调控Th17细胞因子IL-22、IL-17表达和该模型疾病的病理进程。     

IL-33对人永生化角质形成细胞系和模型小鼠银屑病样皮损的影响

目的观察白细胞介素-33(IL-33)对人永生化角质形成细胞系(HaCAT)和咪喹莫特(IMQ)诱导的小鼠银屑病样皮损的影响,并对其可能的机制进行探讨。方法不同浓度IL-33刺激HaCAT细胞;CCK8法检测细胞增殖;Western blot检测细胞中LC3、Beclin1和p-STAT3的表达。将BALB/c雌性小鼠随机分为:正常对照组、IMQ组(背部外涂5%咪喹莫特乳膏,建立银屑病样皮损模型)和IL-33处理组(腹腔注射IL-33)。Western blot检测皮损组织中自噬相关蛋白的表达。结果与对照组相比,25 ng/mL的IL-33处理组促进HaCAT细胞的增殖作用最明显(P<0.05),同时,IL-33可促进LC3、Beclin1和p-STAT3蛋白的表达(P<0.05)。在小鼠银屑病样模型中,与正常对照组相比,IMQ组小鼠背部出现鳞屑和红斑伴有表皮增厚,而IL-33处理组与IMQ组相比小鼠背部皮损鳞屑、红斑及增厚更明显(P<0.01)。IL-33处理组皮损中自噬相关蛋白的表达水平低于IMQ组(P<0.05),p-STAT3蛋白的表达水平均高于正常对照组和IMQ组(P<0.05)。结论IL-33促进HaCAT细胞的增殖,诱导细胞自噬,加重咪喹莫特诱导的小鼠银屑病样皮损的发生和发展,其机制可能是通过引起STAT3信号通路的活化而实现的。     

过表达IL-17RLM的人脐带间充质干细胞对TNBS诱导的结肠炎小鼠脾脏淋巴细胞的免疫调节作用

目的:研究过表达白细胞介素17受体样分子的人脐带间充质干细胞(IL-17RLM-hUCMSCs)对三硝基苯磺酸(TNBS)诱导的结肠炎小鼠脾脏淋巴细胞的免疫调节作用,为炎症性肠病的干细胞治疗提供优化的种子细胞。方法:体外分离培养hUCMSCs,利用慢病毒载体向干细胞内转入IL-17RLM基因,构建IL-17RLM-hUCMSCs。采用TNBS诱导小鼠实验性结肠炎模型,无菌取炎症小鼠脾脏制备淋巴细胞悬液,在刀豆蛋白A(Con A)刺激下将淋巴细胞分别与不同浓度的IL-17RLM-hUCMSCs及hUCMSCs共培养,72 h后以淋巴细胞+Con A为阳性对照,CCK8法及CFSE标记法检测淋巴细胞的增殖情况;同时用流式细胞术检测T淋巴细胞亚群(Th1、Th2、Th17及Treg)比例的改变。结果:hUCMSCs及IL-17RLM-hUCMSCs均对ConA刺激下的淋巴细胞增殖有抑制作用(P0.05);当MSCs/淋巴细胞为1∶1~1∶10时,MSCs对淋巴细胞增殖的抑制作用呈现浓度依赖性。在有效浓度范围内,IL-17RLM-hUCMSCs较h UCMSCs抑制作用更强(P0.05)。hUCMSCs及IL-17RLM-h UCMSCs均能够下调结肠炎小鼠脾脏淋巴细胞的Th1和Th17细胞亚群比例,上调Treg细胞亚群比例,但IL-17RLM-h UCMSCs对Th17细胞亚群的抑制作用更显著(P0.05)。结论:IL-17RLM-hUCMSCs呈浓度依赖性地抑制TNBS诱导的结肠炎小鼠脾脏淋巴细胞增殖,且该作用优于h UCMSCs。同时,IL-17RLM-hUCMSCs可调节T细胞亚群的免疫平衡,且抑制Th17细胞亚群作用强于hUCMSCs。     

Th17细胞及其相关细胞因子在幽门螺杆菌感染中的作用研究

目的 探讨Th17细胞及其相关细胞因子在幽门螺杆菌(H.pylri)感染中的作用.方法 建立幽门螺杆菌感染的小鼠模型,同时设立治疗组与对照组.HE染色评价小鼠胃组织学改变;RT-PCR法检测胃组织IL-17 mRNA、IL-23 mRNA表达水平;ELISA法检测胃组织匀浆上清IL-17、IL-23含量;FCM检测脾脏单细胞悬液中Th17细胞应答情况.结果 与对照组相比,H.pylori感染组小鼠胃组织IL-17、IL-23在mRNA及蛋白水平表达均升高,脾淋巴细胞中Th17细胞比例显著升高;而且H.pylori感染组小鼠IL-17、IL-23 mRNA和蛋白含量以及脾淋巴细胞中Th17细胞比例随感染时间延长而增加;治疗组小鼠IL-17、IL-23表达量及脾淋巴细胞中Th17细胞比率,与治疗前即感染后4周的小鼠相比较均有所下降;感染后不同时期小鼠胃黏膜的炎症程度与IL-17、IL-23的表达量存在正相关.结论 H.pylori感染后可以诱导Th17细胞应答且IL-17、IL-23表达均上调;H.pylori感染后胃炎程度与胃组织1L-17、IL-23含量存在正相关.     

Th17/Treg及其相关细胞因子在结肠癌中的表达并与肿瘤分期的相关性

研究结肠癌患者肿瘤组织中Th17细胞、Treg细胞及患者外周血中相关细胞因子的表达水平,探讨其表达与肿瘤分期的相关性及可能机制。运用流式细胞分析(FACS)技术检测30例结肠癌肿瘤组织及癌旁正常组织中Th17细胞及Treg细胞的比例;采用逆转录聚合酶链反应技术(RT-PCR)检测20例结肠癌患者外周血中Th17、Treg相关细胞因子IL-23和IL-10的表达水平。结果显示结肠癌肿瘤组织中Th17细胞和Treg细胞的比例明显高于癌旁正常组织(P<0.05),进展期肿瘤组织中Treg细胞的比例高于早期(P<0.05),而Th17细胞的比例较早期无明显差异(P>0.05),进展期肿瘤组织中Th17/Treg细胞的比例比早期偏低(P<0.05)。结肠癌患者外周血中IL-23、IL-10的mRNA水平升高,与健康对照组差异明显(P<0.05),且进展期与早期结肠癌IL-10mRNA的表达水平差异显著(P<0.05),而IL-23mRNA在两组间无明显差异(P>0.05)。随着结肠癌病程的进展,肿瘤组织内Th17细胞及Treg细胞的比例逐渐升高,且Treg细胞比Th17细胞升高更加明显。相关细胞因子IL-23和IL-10在患者外周血中的变化趋势和Th17、Treg细胞在肿瘤组织中的变化趋势相一致,提示Th17、Treg细胞在结肠癌的表达可能与肿瘤免疫微环境中相关的细胞因子调节有关。     

Slit2在小鼠银屑病样皮炎模型炎性浸润中的作用

目的 观察分泌型糖蛋白Slit2在咪喹莫特诱导的小鼠银屑病样皮炎模型中的表达及作用。方法 采用免疫荧光观察咪喹莫特诱导的小鼠银屑病样皮损中Slit2的定位,PCR及Western blot检测皮损中Slit2的表达。30只小鼠随机分为5组：正常组、IMQ模型组、Slit2 siRNA组、Slit2蛋白低剂量组（100 ng/ml）、Slit2蛋白高剂量组（300 ng/ml）,每组6只小鼠。每天对小鼠皮肤进行PASI评分,HE染色测量表皮厚度,计数各组真皮内炎症细胞数及血管密度,RT-PCR检测各组皮损处IL-23、IL-17A、IL-22 mRNA表达量,免疫组化检测各组皮损处Ki67表达及CD3 T细胞数量。结果 咪喹莫特诱导的小鼠银屑病样皮炎模型皮损处Slit2的表达升高。相较于模型组,Slit2 siRNA处理后PASI评分增加,表皮变厚、表皮细胞增殖程度增加,真皮炎性细胞浸润增多及血管密度升高,IL-23、IL-17A、IL-22 mRNA水平升高,CD3 T细胞数量明显增加。给予重组Slit2蛋白处理后,PASI评分降低,表皮变薄,表皮细胞增殖程度降低,真皮炎性细胞浸润及血...     

Interleukin-33 alleviates psoriatic inflammation by suppressing the T helper type 17 immune response

Psoriasis is a chronic inflammatory skin disease with unclear pathogenesis. Interleukin-33 (IL-33) is highly expressed in patients with psoriasis, but its role in psoriasis is unknown. The aim of this study was to investigate the possible role of IL-33 in the pathogenesis and treatment of psoriasis. IL-33 expression was determined using enzyme-linked immunosorbent assay, real-time fluorescent quantitative polymerase chain reaction and immunohistochemical staining. CD4⁺ T cells were sorted using magnetic beads and treated with or without IL-33. Imiquimod (IMQ) was used to induce psoriatic inflammation in mice. The frequency of immune cells was determined using flow cytometry. The cytokine level in mouse skin was measured using cytometric bead array. Our results showed that IL-33 was highly expressed in the lesional skin and serum of patients with moderate-to-severe plaque psoriasis. IL-33 inhibited the expression of IL-17 in CD4⁺ T cells of psoriasis patients. Subcutaneous injection of IL-33 alleviated the IMQ-induced psoriatic inflammation in mice, reduced tumor necrosis factor-α and IL-23 expression, and decreased the proportion of T helper type 17 (Th17) cells in the skin-draining lymph nodes in the mice. Our results suggest that IL-33 plays a protective role in the pathogenesis of psoriasis by suppressing Th17 cell differentiation and function. The potential therapeutic effect of IL-33 in treating psoriasis warrants further investigation.     

A comparison of the effects of topical treatment of calcipotriol,camptothecin, clobetasol and tazarotene on an imiquimod-induced psoriasis-like mouse model

The interleukin-23/interleukin 17A (IL-23/IL-17A) cytokine axis plays a critical role in the pathogenesis of psoriasis. In this study, we report the effects of topical calcipotriol, camptothecin, clobetasol and tazarotene on the treatment of imiquimod (IMQ)-induced psoriasis-like inflammation, the development of which is dependent on the IL-23/IL-17A axis. IMQ-induced epidermal hyperplasia and inflammation in the BALB/c mouse ear were significantly inhibited following clobetasol treatment but not calcipotriol, camptothecin or tazarotene treatments. Real-time polymerase chain reaction showed that the mRNA levels of IL-17A, IL-17F, IL-22, IL-1β, IL-6 and TNF-α in ear skin were significantly decreased by clobetasol. In addition, we observed that calcipotriol, camptothecin and tazarotene failed to show any inhibitory effects on the IL-23/IL-17A/IL-22 axis. We also found that clobetasol treatment inhibited the proliferation of γδ T cells and C-C chemokine receptor type 6 (CCR6) expression induced by IMQ. Calcipotriol, camptothecin and tazarotene not only failed to inhibit this proliferation but also enhanced retinoic acid-related orphan receptor γ (RORγ) expression in IMQ-induced psoriasis-like inflammation. In conclusion, we suggest that clobetasol induces the relief of IMQ-induced psoriasis-like inflammation in a mouse model but that calcipotriol, camptothecin and tazarotene cannot. Therefore, we suggest that more in-depth studies on pharmacological effects of tazarotene, camptothecin and calcipotriol should be carried out.     

Unexpected connections of the IL-23/IL-17 and IL-4/IL-13 cytokine axes in inflammatory arthritis and enthesitis

The IL-23/IL-17 cytokine axis is related to spondyloarthropathy (SpA) pattern diseases that target the skin, eye, gut and joints. These share overlapping target tissues with Th2 type or allergic diseases, including the skin, eye and gut but SpA diseases exhibit distinct microanatomical topography, molecular characteristics, and clinical features including uveitis, psoriasis, apical pulmonary involvement, lower gastrointestinal involvement with colitis, and related arthritides including psoriatic arthritis and ankylosing spondylitis. Inflammatory arthritis is conspicuously absent from the Th2 diseases which are characterised IL-4/IL-13 dependent pathway activation including allergic rhino-conjunctivitis, atopic eczema, allergic asthma and food allergies. This traditional understanding of non-overlap of musculoskeletal territory between that atopic diseases and the IL-17 -mediated SpA diseases is undergoing a critical reappraisal with the recent demonstration of IL-4/IL-13 blockade, may be associated with the development of SpA pattern arthritis, psoriasiform skin disease and occasional anterior uveitis. Given the known plasticity within Th paradigm pathways, these findings suggest dynamic Th2 cytokine and Th17 cytokine counter regulation in vivo in humans. Unexpected, this is the case in peripheral enthesis and when the IL-4/13 immunological brake on IL-23/17 cytokines is removed, a SpA phenotype may emerge. We discuss hitherto unexpected observations in SpA, showing counter regulation between the Th17 and Th2 pathways at sites including the entheses that collectively indicate that the emergent reverse translational therapeutic data is more than coincidental and offers new insights into the “Th paradigms” in atopy and SpA.     

Psoriatic inflammation causes hepatic inflammation with concomitant dysregulation in hepatic metabolism via IL-17A/IL-17 receptor signaling in a murine model

Psoriatic inflammation has been shown to be associated with cardiovascular dysfunction and systemic inflammation. Recently, psoriasis has also been linked to hepatic disorders, however underlying mechanism connecting the two are unknown. IL-17A being a central pro-inflammatory cytokine in the pathogenesis of psoriasis may be involved in hepatic inflammation through its receptor and downward signaling; however so far no study has investigated IL-17A related signaling in the liver during psoriasis in a murine model. Therefore, this study explored psoriasis-induced hepatic inflammation and concurrent metabolic changes. Mice were applied topically imiquimod (IMQ) to develop psoriatic inflammation. Additionally mice were also treated either with IL-17A or anti-IL17A antibody to explore the role of IL-17 related signaling in liver. Mice were then assessed for hepatic inflammation through assessment of inflammatory/oxidative stress markers (IL-17RC, NFκB, IL-6, MCP-1, IL-1β, GM-CSF, ICAM-1, iNOS, lipid peroxides and myeloperoxidase activity) as well as hepatic injury (alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase) and protein/lipid metabolic biomarkers (total proteins, albumin, total bilirubin, triglycerides, HDL cholesterol, and total cholesterol). IMQ treatment led to hepatic inflammation as evidenced by increased pro-inflammatory cytokines and oxidative stress with concomitant dysregulation in hepatic protein/lipid metabolism. Treatment with IL-17A further aggravated, whereas treatment with anti-IL17A antibody ameliorated IMQ-induced changes in hepatic injury/inflammation and protein/lipid metabolism. Our study shows for the first time that psoriatic inflammation leads to hepatic inflammation which results in dysregulated protein/lipid metabolism through IL-17RC/NFκB signaling. This could result in increased risk of cardiovascular dysfunction in patients with psoriasis.     

The CCL20 and CCR6 axis in psoriasis

Psoriasis is a TNF-α/IL-23/IL-17A–mediated inflammatory skin disease that causes a significant socioeconomic burden in afflicted patients. IL-17A–producing immune cells, including Th17 cells, are crucial effector cells in the development of psoriasis. IL-17A stimulates epidermal keratinocytes to produce CCL20, which eventually recruits CCR6 + Th17 cells into the lesional skin. Thus, the CCL20/CCR6 axis works as a driving force that prepares an IL-17A–rich cutaneous milieu. In this review, we summarize the current research topics on the CCL20/CCR6 axis and the therapeutic intervention of this axis for psoriasis.     

The Immunologic Role of IL-17 in Psoriasis and Psoriatic Arthritis Pathogenesis

Psoriasis is a chronic, immune-mediated, inflammatory disease that is pathogenically driven by proinflammatory cytokines. This article reviews the immunologic role of interleukin (IL)-17, the major effector cytokine in the pathogenesis of psoriatic disease, along with the rationale for targeting the IL-17 cytokine family (IL-17A, IL-17F, and IL-17 receptor A) in the treatment of psoriasis and psoriatic arthritis. Emerging evidence indicates that major sources of IL-17A in patients with psoriatic disease are mast cells, γδ T cells, αβ T cells, and innate lymphoid cells in lesional skin and synovial fluid. Within the skin and joints, IL-17A acts on cellular targets, including keratinocytes, neutrophils, endothelial cells, fibroblasts, osteoclasts, chondrocytes, and osteoblasts, to stimulate production of various antimicrobial peptides, chemokines, and proinflammatory and proliferative cytokines, which, in turn, promote tissue inflammation and bone remodeling. The critical importance of the IL-23/IL-17A axis to the pathogenesis of psoriatic disease has resulted in many new biologic treatments targeting these cytokines. These biologics dramatically improve skin and joint symptoms in patients with moderate-to-severe psoriasis and psoriatic arthritis.     

银屑病发病机制中IL-17关键轴与IgE的相互作用

银屑病是一种免疫介导的多基因遗传性皮肤病,其发病机制尚未明确,目前认为可能是遗传、环境和免疫学因素共同作用的结果,在免疫机制方面,白细胞介素(interleukin,IL)-23 /辅助T细胞(helper T cells,Th)17途径已被确定为关键轴,促炎细胞因子IL-17A是其主要作用因子。有研究表明,银屑病患者血清中免疫球蛋白(immune globulin,Ig)E浓度增加。与非病变皮肤相比,病变皮肤有更多的特异性IgE的Fc段高亲和力受体I (receptor I for the Fc region of IgE,FceRI)相关细胞,包括肥大细胞,表皮树突状细胞,朗格汉斯细胞和巨噬细胞,现就银屑病发病机制中IL-17关键轴与IgE的相互作用进行综述。     

IL-2协同抗CD45RB抗体调节Treg/Th17细胞的比例并延长小鼠移植皮肤存活时间

目的:研究IL-2在抗CD45RB抗体诱导免疫耐受中对Treg/Th17细胞分化的影响,进一步阐明抗CD45RB抗体诱导免疫耐受的机制。方法:用免疫磁珠分选C57BL/6小鼠脾脏中的CD4+T细胞,在抗CD45RB抗体与IL-2的作用下培养72小时后,流式检测Treg/Th17细胞的变化。以BALB/c小鼠为供体,C57BL/6小鼠为受体建立同种异基因皮肤移植模型,分别给予抗CD45RB抗体及IL-2等治疗,术后1、3、5、7、9天取受体鼠脾细胞,动态检测Treg/Th17细胞的变化;术后第9天取移植皮肤HE染色观察炎性细胞的浸润情况;观察并记录移植皮肤的存活时间。结果:CD4+T细胞在IL-2联合抗CD45RB抗体的作用下培养72小时后,Treg比例升高,Th17细胞比例下降;IL-2联合抗CD45RB抗体治疗后明显延长小鼠移植皮肤的存活时间。结论:IL-2可以明显增强抗CD45RB抗体诱导免疫耐受的形成,使Treg细胞上调,下调Th17细胞,有利于免疫耐受的形成。