地塞米松对哮喘大鼠Th1/Th2失衡及嗜酸性粒细胞凋亡的作用

目的　观察地塞米松对哮喘大鼠Th1/Th2 (T辅助细胞 1/T辅助细胞 2 )平衡、嗜酸性粒细胞 (EOS)凋亡率的影响作用 ,探讨糖皮质激素治疗哮喘的机制。方法　清洁级雄性SD大鼠 30只 ,随机分为 3组 :正常对照组 (C)、哮喘组 (A)、地塞米松治疗组 (T)。卵清白蛋白复制大鼠哮喘模型 ,检测大鼠血、支气管肺泡灌洗液 (BALF)中IL 4、IFN γ水平及气道壁EOS凋亡率。结果　A组大鼠BALF中IFN γ水平明显低于C组 (P <0 0 1) ,血中及BALF中的IL 4水平则明显高于C组 (均为P<0 0 1) ,表现出明显的Th1/Th2失衡。T组大鼠IL 4的表达明显低于A组而IFN γ的表达则明显高于A组 (均为P <0 0 1)。在A组 ,大鼠气道壁EOS的浸润明显增多 ,但凋亡细胞却很少看到 ,EOS凋亡率明显低于C组 (P <0 0 1) ,T组EOS凋亡率则明显高于A组 (P <0 0 1)。结论　纠正哮喘Th1/Th2失衡和促进气道壁EOS凋亡可能是糖皮质激素减轻哮喘气道炎症的重要机制之一。     

支气管哮喘与Th1/Th2/Th17的相关性分析

支气管哮喘是一种涉及多种临床症状,例如咳嗽、气喘、呼吸困难等常见的呼吸系统疾病,其发病率之高严重威胁着人类的健康.哮喘的发病与遗传和环境都有着密切的联系,因其发病机制十分复杂至今尚未明了.众多的细胞及其细胞因子均参与了哮喘的发病过程,其中辅助性T细胞(Thelper,Th)及其亚群(Th1、Th2和Th17)已被证实与支气管哮喘的发生有着十分密切的关系.     

枯草芽孢杆菌ATCC9372 产生海藻酸寡糖通过T 辅助细胞2 型相关细胞因子能有效抑制哮喘发作的研究

目的:研究海藻酸寡糖(AO)通过T辅助细胞2型相关细胞因子治疗哮喘疾病的疗效。方法:8周龄的SPF级小鼠共32只,雌雄各半,随机分为4组,分别为正常对照组(n=8)、哮喘模型组(n=8)、海藻酸寡糖50 mg/kg组(n=8)和海藻酸寡糖200 mg/kg组(n=8)。在卵清蛋白(OVA)诱导的小鼠哮喘模型中,给予AO两种不同的剂量,分别为50 mg/kg和200 mg/kg。在支气管肺泡灌洗液(BALF)中,通过免疫组化和RT-PCR等方法,分析和比较治疗组和模型组两组的T辅助细胞2型相关细胞因子表达的差异,以及两种剂量的组间差异。结果:与正常组比较,哮喘组BALF中炎性细胞和嗜酸性粒细胞、血清总IgE、BALF中Th2和Th1相关细胞因子的表达(CD3、CD4、CD8和CD68)以及mRNA表达明显增高(P0. 05)。与哮喘组比较,AO治疗组各项指标均明显降低(P0. 05),而且高、低剂量组之间差异有统计学意义(P0. 05)。结论:由于海藻酸寡糖调节Th1/Th2细胞因子是导致哮喘发病的机制。因此,AO治疗哮喘疾病的前景十分可观,可以成为治疗哮喘的候选药物。     

哮喘急性发作期患儿外周血Th17水平及其功能初步研究

目的:观察哮喘急性发作期患儿外周血Th17水平和功能状态,初步探讨Th17在儿童哮喘发病中的作用.方法:以正常儿童(对照组,n=20)为对照,流式细胞术(FCM)检测哮喘急性发作期患儿(哮喘组,n=26)外周血Th17以及Th1,Th2细胞百分率,ELISA方法检测外周血单个核细胞(PBMC)体外培养上清IL-17,IFN-γ以及血浆IL-17,IL-23,IgE水平.结果:(1) 哮喘组和对照组外周血CD4+T中CD4+IL-17+T比例[(1.25±0.66)% vs (1.27±0.66)%(P>0.05)];CD4+IL-4+T比例[(2.40±2.55)% vs(2.52±1.26)%(P>0.05)];两组比较无统计学意义.哮喘组CD4+IFN-γ+T比例低于对照组[15.79±7.48 vs 24.10±12.70(P<0.05)].(2) PHA活化的PBMC体外培养72 h上清中,哮喘组IL-17水平为17.53(0～245.57)ng/L,低于对照组101.74(25.12～500.60)ng/L(P<0.01);IFN-γ水平与对照组比较无统计学意义[3507±2788 ng/L vs 3027±2737 ng/L];两组血浆中IL-17均<2 ng/L,IL-23均<15 ng/L.(3) 哮喘组血浆总IgE 499.26(2.3～945.1)IU/mL,高于对照组54.57(1.7～318.26)IU/mL(P<0.001);IgE阳性(>150 IU/mL)和阴性(<150 IU/mL)哮喘患儿体外PBMC培养上清中IL-17,IFN-γ水平比较无统计学意义.结论:哮喘急性发作期患儿外周血Th1细胞以及PHA活化的IL-17表达水平降低,Th1水平和Th17功能异常的机制及其在儿童哮喘发病中的作用值得进一步研究.     

重症支气管哮喘患者CD4T淋巴细胞免疫功能分析

目的探讨重症支气管哮喘患者CD4 T淋巴细胞免疫功能及相关细胞因子的变化。方法选择本院收治的重症支气管哮喘患者70例(实验组)及健康人群70例(对照组)作为研究对象,采用流式细胞仪检测CD4+T细胞亚群Th1、Th2、Th17以及Treg细胞在研究对象外周血中的比例;采用荧光定量PCR检测研究对象外周血PBMC细胞IFN-γ、IL-2、IL-4、IL-10、IL-17、IL-22、TGF-β、IL-10的mRNA表达水平;采用ELISA法检测研究对象血清中IFN-γ、IL-2、IL-4、IL-17、IL-22、TGF-β、IL-10的蛋白表达浓度。结果与对照组相比,实验组外周血中Th1/Th2细胞比例显著下降,而Th17/Treg细胞比例显著增高(P0.05);实验组外周血PBMC细胞中IL-4、IL-10、IL-17、IL-22的m RNA、蛋白表达水平均显著上调(P0.05);而IFN-γ、IL-2、TGF-β的mRNA、蛋白表达水平则呈现明显下降(P0.05)。结论重症支气管哮喘患者存在Th1/Th2、Th17/Treg的细胞免疫平衡失调,这种细胞免疫应答的变化可能与重症支气管哮喘的疾病进展密切相关。     

热休克蛋白90调控抗原交叉提呈及其在支气管哮喘中的作用

树突状细胞(DC)是机体功能最强的专职抗原提呈细胞(APC).DC在启动、调控和维持免疫应答中发挥中心作用,促进初始型T细胞(Th0细胞)向不同的T细胞亚群分化.热休克蛋白是生物体中普遍存在和高度保守的蛋白质.表达于DC的热休克蛋白90(HSP90)在DC抗原提呈中起重要作用.支气管哮喘(简称哮喘)是多基因参与的具有遗传易感性的慢性气道炎症性疾病,其发病涉及多种炎症细胞、炎性介质和复杂的细胞因子网络,以血清IgE增高、肺组织嗜酸性粒细胞浸润和气道高反应性为显著的临床特征[1].现认为,哮喘系DC介导的Ⅱ型辅助性T细胞(Th2)优势免疫为特征的慢性气道变应性疾病[2].现就HSP90的生物学特性、DC交叉抗原提呈及其与哮喘关系的研究进展做一综述.     

过敏性哮喘患者调节性T细胞对Th17细胞和Th9细胞的影响

目的:深入研究过敏性哮喘病人(轻到中度)急性发作期的调节性T细胞对Th17和Th9细胞功能影响的异常。方法:招募30例过敏性哮喘病人(轻到中度)急性发作期和30例健康者,采集外周静脉血30 ml,分离PBMC,然后应用磁珠法体外分离CD4+CD25+CD127-/low调节性T细胞(Tregs)和CD4+CD25-效应T细胞(选取分离纯度90%以上的标本继续下一步试验)。在PHA刺激下体外培养,检测效应T细胞增殖情况和Th17细胞、Th9细胞特异性基因(RORC和PU.1)表达及特异性细胞因子(IL-17和IL-9)分泌情况,检测哮喘病人Th17细胞和Th9细胞的异常情况;并给予Tregs干预,检测Tregs对Th17细胞和Th9细胞分化的影响。结果:哮喘组和健康对照组的Tregs均可以抑制CD4+CD25-效应细胞增殖反应,但哮喘组Tregs的抑制功能较健康对照组明显下降(P=0.03)。哮喘病人RORC表达和IL-17分泌均高于对照组(P0.05),Tregs对RORC表达的抑制能力在哮喘组有所下降(P0.05),但对IL-17分泌的抑制能力在哮喘组和对照组没有显著差别(P0.05);哮喘病人IL-9分泌无论是否加入Tregs干预均高于对照组(P0.05),但PU.1表达只有在加入Tregs干预后才高于对照组(P=0.04);Tregs对PU.1表达和IL-9分泌的抑制能力在哮喘组和对照组没有显著差别(P0.05)。结论:过敏性哮喘病人(轻到中度)急性发作期Tregs数量和功能下降,而Th17细胞和Th9细胞的特异性基因表达和特异性细胞因子生成增加,哮喘病人的Tregs在体外对Th17细胞的抑制功能有所下降,但尚未发现Tregs对Th9细胞的抑制功能明显降低。     

扁蒴藤素对支气管哮喘患儿Th17 / Treg 失衡的影响及意义

目的:探讨扁蒴藤素对支气管哮喘患儿Th17/ Treg 体外培养细胞的影响及意义,从而探讨免疫因素在支气管哮喘治疗中的机理。方法:提取支气管哮喘患儿外周血单个核细胞,并在体外给予扁蒴藤素干预,流式细胞术检测Th17 及Treg 比例,ELISA 方法检测细胞培养上清液中IL-17 及TGF-β含量变化,实时定量PCR 检测ROR-T 及Foxp3 含量。结果:扁蒴藤素干预组与对照组相比,CD4+ IL-17+ T 细胞(即Th17 细胞)表达量显著降低,CD4+ CD25+ Foxp3+ T 细胞(即Treg 细胞)表达量显著升高,各组均存在统计学意义(P<0.05);与对照组相比,IL-17 含量显著降低,TGF-β含量显著升高,各组均存在统计学意义(P<0.05);与对照组相比,ROR T 含量显著降低,Foxp3 含量显著升高,各组均存在统计学意义(P<0.05)。结论:扁蒴藤素干预可较大程度缓解体外培养Th17/ Treg 细胞失衡,提示该药物可能通过影响免疫平衡来调节哮喘的发生发展,为扁蒴藤素在临床治疗哮喘的应用提供了理论支持。     

气道上皮细胞在过敏性哮喘固有免疫应答中的作用研究进展北大核心CSCD

过敏性哮喘是一种发病率逐年升高的慢性疾病,典型表现是气道炎症和气道高反应性。气道上皮细胞(AEC)作为抵御多种过敏原的第一道屏障,能够率先启动固有免疫应答参与过敏性哮喘,AEC通过分泌抗微生物物质、表达模式识别受体、分泌固有免疫因子、参与固有免疫记忆的方式调节适应性免疫应答,影响过敏性哮喘的进程。因此AEC被认为是过敏性哮喘中肺部炎症和免疫应答的调节枢纽。另外,针对AEC固有免疫功能的靶向治疗药物也得到了快速发展,为临床治疗过敏性哮喘提供了更多选择。     

哮喘小鼠中核因子-KB对Th2型细胞因子分泌的影响

支气管哮喘表现为Th1／Th2型细胞免疫应答的平衡失调、Th1型细胞的免疫应答下调、Th2型细胞因子(如IL-4、IL-5等)过度表达。核因子-KB(NF—KB)作为一种具有广泛生物学活性的核转录因子，参与了许多炎症因子表达的调控：有资料提示哮喘患者的T细胞中NF—KB激活后可增加IL-4、IL-5基因的转录，使IL-4、IL-5的表达增加，而导致哮喘炎症     

Immunopathogenesis of Allergic Asthma: More Than the Th2 Hypothesis

Asthma is a chronic obstructive airway disease that involves inflammation of the respiratory tract. Biological contaminants in indoor air can induce innate and adaptive immune responses and inflammation, resulting in asthma pathology. Epidemiologic surveys indicate that the prevalence of asthma is higher in developed countries than in developing countries. The prevalence of asthma in Korea has increased during the last several decades. This increase may be related to changes in housing styles, which result in increased levels of indoor biological contaminants, such as house dust mite-derived allergens and bacterial products such as endotoxin. Different types of inflammation are observed in those suffering from mild-to-moderate asthma compared to those experiencing severe asthma, involving markedly different patterns of inflammatory cells and mediators. As described in this review, these inflammatory profiles are largely determined by the involvement of different T helper cell subsets, which orchestrate the recruitment and activation of inflammatory cells. It is becoming clear that T helper cells other than Th2 cells are involved in the pathogenesis of asthma; specifically, both Th1 and Th17 cells are crucial for the development of neutrophilic inflammation in the airways, which is related to corticosteroid resistance. Development of therapeutics that suppress these immune and inflammatory cells may provide useful asthma treatments in the future.     

Emerging Roles of T Helper Subsets in the Pathogenesis of Asthma

The cardinal features of asthma include pulmonary inflammation and airway hyperresponsiveness (AHR). Classically, asthma, specifically allergic asthma, has been attributed to a hyperactive Th2 cell immune response. However, the Th2 cell-mediated inflammation model has failed to adequately explain many of the clinical and molecular aspects of asthma. In addition, the outcomes of Th2-targeted therapeutic trials have been disappointing. Thus, asthma is now believed to be a complex and heterogeneous disorder, with several molecular mechanisms underlying the airway inflammation and AHR that is associated with asthma. The original classification of Th1 and Th2 pathways has recently been expanded to include additional effector Th cell subsets. These include Th17, Th9 and Treg cells. Emerging data highlight the involvement of these new Th cell subsets in the initiation and augmentation of airway inflammation and asthmatic responses. We now review the roles of these recently classified effector Th cell subsets in asthmatic inflammation and the insights they may provide in addition to the traditional Th2 paradigm. The hope is that a clearer understanding of the inflammatory pathways involved and the mediators of inflammation will yield better targeted therapeutics.     

The sentinel role of the airway epithelium in asthma pathogenesis

The adoption of the concept that asthma is primarily a disease most frequently associated with elaboration of T-helper 2 (Th2)-type inflammation has led to the widely held concept that its origins, exacerbation, and persistence are allergen driven. Taking aside the asthma that is expressed in non-allergic individuals leaves the great proportion of asthma that is associated with allergy (or atopy) and that often has its onset in early childhood. Evidence is presented that asthma is primarily an epithelial disorder and that its origin as well as its clinical manifestations have more to do with altered epithelial physical and functional barrier properties than being purely linked to allergic pathways. In genetically susceptible individuals, impaired epithelial barrier function renders the airways vulnerable to early life virus infection, and this in turn provides the stimulus to prime immature dendritic cells toward directing a Th2 response and local allergen sensitization. Continued epithelial susceptibility to environmental insults such as viral, allergen, and pollutant exposure and impaired repair responses leads to asthma persistence and provides the mediator and growth factor microenvironment for persistence of inflammation and airway wall remodeling. Increased deposition of matrix in the epithelial lamina reticularis provides evidence for ongoing epithelial barrier dysfunction, while physical distortion of the epithelium consequent upon repeated bronchoconstriction provides additional stimuli for remodeling. This latter response initially serves a protective function but, if exaggerated, may lead to fixed airflow obstruction associated with more severe and chronic disease. Dual pathways in the origins, persistence, and progression of asthma help explain why anti-inflammatory treatments fail to influence the natural history of asthma in childhood and only partially does so in chronic severe disease. Positioning the airway epithelium as fundamental to the origins and persistence of asthma provides a rationale for pursuit of therapeutics that increase the resistance of the airways to environmental insults rather than concentrating all effort on suppressing inflammation.     

Pathophysiology of severe asthma

Although asthma affects nearly 8% of the adult population, most of these patients have mild-to-moderate disease that can be controlled with appropriate treatment. It is estimated, however, that 5% to 10% of patients with asthma have severe disease that is unresponsive to typical therapeutics, including corticosteroids. Because patients with severe asthma are disproportionately affected by their disease, in terms of both impaired lifestyle and health care costs, the National Heart, Lung, and Blood Institute sponsored a workshop on the pathogenesis of severe asthma. The goals of this workshop were to begin to define the characteristics of severe asthma. In these discussions, it was clear that many characteristics need to be considered in defining this phenotype of asthma, including symptoms, intensity of therapy (including administration of systemic corticosteroids), and impairment of lung function. Also discussed were potential mechanisms of severe asthma including the role of allergic diseases, which may play less of a role in severe asthma than in mild-to-moderate disease, and infections. A major limitation to control of severe asthma is the recalcitrant response of these patients to usual therapy including systemic corticosteroids; the potential of other therapies was reviewed. From these discussions, recommendations were made for future research needs to gain insights into a difficult therapeutic and possibly novel mechanistic area of asthma.     

Anti-TNF-alpha and Th1 cytokine-directed therapies for the treatment of asthma

PURPOSE OF REVIEW: This article examines recent work about the role of TNF-alpha and of selected Th1-related cytokines in asthma with particular emphasis on the therapeutic potential of blocking the biological activity of these mediators. RECENT FINDINGS: Current research endeavours suggest that asthma pathogenesis is driven by a mixed Th1/Th2 immune response. The contribution of individual Th1-associated and Th2-associated effector mechanisms to this mixed response profile is highly heterogeneous and variations in response patterns seem to be associated with heterogeneity in clinical phenotypes. In particular, it is now acknowledged that allergen-specific Th1 responses appear to be responsible for the pathogenetic effects seen in patients suffering from the more severe chronic forms of the disease. This is important because usual treatments for asthma appear to have limited effects on the more chronic severe forms of the disease and there is a pressing need for the development of new treatment strategies. The failure of topical corticosteroids to reduce TNF-alpha and Th1-derived cytokines to a significant level in asthmatic airways may explain to a certain extent why these drugs appear to have limited effects in the more severe forms of asthma. SUMMARY: It is likely that therapies blocking TNF-alpha and interfering with Th1-derived cytokines may be a considerable advance in the management of those asthma patients who are particularly resistant to typical treatment modalities.     

Neutrophilic inflammation in severe asthma

Neutrophils may play an important role in the pathogenesis of severe asthma. Their infiltration into the airway is increased. Interleukin (IL)-8 is involved in this process, and is actually upregulated in the airways of patients. We have observed that in the absence of eosinophil chemoattractants, neutrophils stimulated by IL-8 augment eosinophil trans-basement membrane migration by releasing superoxide anion, matrix metalloproteinase, leukotriene B(4) and platelet-activating factor. These findings suggest that IL-8-stimulated neutrophils could lead eosinophils to accumulate in the airways of asthmatic patients, which might be a mechanism for corticosteroid resistance in severe asthma. However, the mechanisms of IL-8 upregulation in the airway are not completely understood. Several studies suggest that IL-17 (or T helper 17 cells; Th17) is involved in the IL-8 upregulation observed in severe asthma. We clarified that dopamine induces Th17 differentiation through dopamine D1-like receptor (D1-like-R), and that the D1-like-R antagonist attenuates Th17-mediated diseases like experimental autoimmune encephalomyelitis. Furthermore, we demonstrated that a D1-like-R antagonist significantly suppressed ovalbumin (OVA)-induced neutrophilic airway inflammation in OVA T cell receptor-transgenic DO11.10 mice through inhibiting Th17-mediated immune responses. Therefore, dopamine D1-like-R antagonists could become useful for treating Th17-mediated neutrophil-dominant severe asthma. As inhaled corticosteroids are known to be less effective for controlling neutrophilic inflammation, a more effective therapeutic strategy for neutrophil-dominant asthma should still be elucidated.     

支气管哮喘患者外周血Th17 与Th9 细胞的变化及临床意义

目的:探讨Th17、Th9 细胞水平在支气管哮喘患者中的表达及其临床意义。方法:选取67 例哮喘患者为研究对象,30 例健康体检者为对照组,根据全球哮喘防治倡议(GINA)将哮喘患者分为间歇与轻度持续组20 例,中度持续组25 例和重度持续组22 例。收集患者临床资料,完成哮喘控制测试(ACT)评分、肺功能和呼出气一氧化氮(FeNO)值测定,流式细胞仪检测外周血PBMC 中CD3+ CD8- IL-17+ 、CD3+ CD8- IL-9+细胞水平。结果:与健康对照组相比,哮喘患者Th17、Th9 细胞水平均明显升高(P<0.05);中重度哮喘患者Th17 及Th9 细胞水平显著高于间歇与轻度哮喘组(P<0.05),而间歇与轻度哮喘组患者与健康对照组比较,差异无统计学意义(P>0.05)。哮喘患者外周血Th17 及Th9 细胞的表达均与FeNO 值正相关(r 值依次为0.501、0.438;P<0.05),与ACT 评分、肺功能FEV1、FEV1/ FVC 及PEF 负相关(r 值依次为-0.441、-0.362;-0.307、-0.377;-0.419、-0.411;-0.428、-0.24;P<0.05),同时Th17 与Th9 细胞在哮喘患者外周血中的表达率正相关(r 值为0.443,P<0.05)。结论:Th17、Th9 细胞在哮喘患者中高表达,具有协同致炎作用,且与症状严重程度相关,提示Th17 及Th9 细胞对哮喘患者病情评估有潜在的参考价值。     

过敏性哮喘患者外周血CD4+T细胞中miR-155的表达及临床意义

目的:探讨过敏性哮喘患者外周血CD4+T细胞中miR-155及其前体基因BIC的表达及临床意义.方法:应用实时荧光定量聚合酶链反应检测不同严重程度哮喘患者50例及健康对照20例外周血CD4+T细胞中BIC及miR-155的表达水平,并且与哮喘患者临床指标进行相关性分析;运用过表达或抑制,探讨miR-155在CD4+T细胞体外分化中的作用.结果:哮喘患者组BIC及miR-155的表达较对照组显著降低(P＜0.01),中、重度哮喘患者的BIC表达均低于对照组(P＜0.05),轻、中、重度哮喘患者的miR-155表达显著低于对照组(P＜0.01);组间比较发现,轻、中、重度哮喘患者的BIC表达无统计学意义,而重度哮喘患者miR-155的表达较轻度哮喘组有明显下降(P ＜0.05);miR-155的表达与用力呼气第1秒量之间呈正相关(P＜0.01);miR-155过表达促进CD4+T细胞向Th1分化,而抑制miR-155的表达则促进CD4+T细胞向Th2分化.结论:过敏性哮喘患者外周血CD4+T细胞中miR-155表达下调,其表达水平与哮喘严重程度相关,提示miR-155调控CD4+T细胞分化在哮喘中发挥重要作用.     

What goes up must come down: biomarkers and novel biologicals in severe asthma

Asthma is a heterogeneous airway disease characterized by typical symptoms in combination with variable airway obstruction. Most patients with asthma have well controlled symptoms and a low risk of asthma attacks with inhaled corticosteroid (ICS) treatment. However, a clinically important subgroup (~ 10%) remains symptomatic and/or at risk of asthma attacks despite maximum inhaled therapy. Patients with severe asthma are responsible for a significant proportion of healthcare costs attributable to asthma and have a large unmet need for better treatments. An important advance in recent years has been the recognition that severe asthma is heterogeneous with respect to clinical problems and the pattern of lower airway inflammation. Identification of eosinophilic inflammation in the airways has become an important priority as novel biologicals that target Th2 cytokines, such as anti‐IL5, anti‐IL‐13 and combined anti‐IL‐4/13 are showing considerable promise as treatments for this subgroup. It has also become clear that anti‐IgE (Omalizumab), the first monoclonal antibody registered for treatment of severe asthma, is only active in patients with active eosinophilic airway inflammation. The future will be identification of potentially responsive patients on the basis of raised biomarkers and, as suggested by the title of this review, targeted treatment with specific cytokine blockade that has a direct effect on the biomarkers. In this review, we outline an approach to the clinical assessment of patients potentially suitable for biological treatment and describe in detail the likely clinical impact of established and new biological treatments.