补脾益气方药对哮喘大鼠Th1/Th2失衡的调节及机制

目的观察补脾益气方药对哮喘大鼠Th1/Th2失衡的免疫调节作用及机制。方法 SPF级雄性Wistar大鼠随机分为对照组、哮喘组、生理盐水治疗组、地塞米松组、补脾益气方药治疗组。检测支气管肺泡灌洗液(BALF)中嗜酸性细胞、巨噬细胞、淋巴细胞和中性粒细胞比例;采用ELASA方法检测BALF中IL-4和IFN-γ的含量;采用Western blot方法检测肺组织GATA-3和T-bet的表达。结果模型组和生理盐水治疗组与正常组比较,BALF中嗜酸性粒细胞、淋巴细胞和中性粒细胞的比例及IL-4含量均明显升高,IFN-γ含量显著下降;GATA-3表达增高,T-bet表达降低。经补脾益气方药与地塞米松治疗后,BALF中嗜酸性粒细胞、淋巴细胞和中性粒细胞的比例和IL-4含量均明显下降,IFN-γ含量显著升高;GATA-3表达下调,T-bet表达上调。上述结果均差异显著,具有统计学意义(P<0.01)。结论补脾益气方药对Th1/Th2失衡有调节作用,这种作用是通过调节GATA-3和T-bet的协调表达来实现。     

白藜芦醇对JurcatT细胞分化为Th1/Th2细胞作用的影响及机制

目的探讨白藜芦醇调节Jurkat T细胞分泌为Th1/Th2型细胞作用的影响。方法将不同浓度的白藜芦醇作用于受植物血凝素（PHA）刺激的Jurkat T细胞,于不同时间点分别用ELISA法检测Th1型细胞因子IFN-γ和Th2型细胞因子IL-10;RT-PCR法检测T-bet和GATA-3 mRNA表达。结果白藜芦醇作用后IFN-γ、IL-10及T-bet mRNA、GATA-3 mRNA的表达均明显降低,具有浓度和时间依赖性。结论白藜芦醇对受PHA刺激的JurcatT细胞由Th0向Th1、Th2细胞分化起抑制作用。     

HIV infection of primary CD4+ Th2 cells, defined by expression of the chemoattractant receptor-homologous (CRTH2), induces a Th0 phenotype

The association between HIV, cytokine profile, and disease progression is controversial. In this study, we evaluated whether HIV infection of a primary T helper-like type 2 cytokine (Th2) cell subset augments their cytokine profile. We utilized the CRTH2 (chemoattractant receptor-homologous) marker to identify CD4+ Th2 cells. Approximately 2-4% of CD4+ T cells are CRTH2+. CRTH2+ expression is confirmed to delineate a Th2 subset as indicated by robust inducible IL-4 response. CD4+ CRTH2+ T cells were also more inherently activated than their CRTH2-negative counterpart as indicated by a higher percent expression of CD69, CD45RO, CD95, CD25, and HLA-DR. CD4+CRTH2+ T cells were not terminally differentiated as indicated by expression of CD27 and CD28. In vitro HIV infection of primary human CD4 CRTH2T cells, independent of chemokine coreceptor usage, potently upregulated IFN-gamma production while still maintaining robust IL-4 expression. This Th0 (IFNgamma+ IL-4+) phenotype was upregulated in CD4+CRTH2+ T cells post-HIV infection by 18-fold, demonstrating a shift to a Th0 phenotype. Ex vivo studies also demonstrated that HIV+ patients exhibited a decline in CD4+CRTH2+ cells and a shift of this population toward cells that express both IFN-gamma and IL-4. Collectively, these data indicate that HIV replication in Th2 cells induces a Th0 phenotype. This phenomenon may be a deliberate viral escape mechanism to prevent the skewing of the immune response toward Th1 or Th2.     

Suppressive oligodeoxynucleotides inhibit atherosclerosis in ApoE(-/-) mice through modulation of Th1/Th2 balance

Atherosclerosis is a chronic inflammatory disease, which is positively and negatively regulated by T helper (Th) 1 and Th2 lymphocytes, respectively. Recent findings indicate that suppressive oligodeoxynucleotides (ODNs) expressing TTAGGG motifs selectively reduce Th1 cytokine production and have been proven effective at blocking the development of organ-specific autoimmune diseases. In the current research, we hypothesized that suppressive ODNs may alter the development of atherosclerosis. Eight-week-old homozygous ApoE^−/− male mice were injected with 300 μg ODNs A151 (TTAGGG) or nonspecific ODNs 1612. Atherosclerotic lesion sizes were dramatically reduced by ODNs A151, but not by nonspecific ODNs. MCP-1 and VCAM-1, which are the key inflammatory factors in atherogenesis, were significantly attenuated by the suppressive ODNs A151. In the splenic lymphocytes, FACS analysis showed ODNs A151 reduced the percentage of IFN-γ-producing Th1 cells and slightly increased the percentage of IL-4-producing Th2 cells, indicating that suppressive ODNs skewed the Th1/Th2 balance toward Th2 inflammation in vivo. Furthermore, ODNs A151 down-regulated the phosphorylation of STAT1 and STAT4 and suppressed up-regulation of T-bet, a signal modulator for Th1, and didn't impact GATA-3 and STAT6, which are associated with a Th2 phenotype. Consistent with this in vivo observation, ELISA analysis demonstrated that ODNs A151 suppressed Th1 cytokines IFN-γ and TNF-α, and augmented Th2 cytokines IL-4 and IL-10 in vitro. This study provides the first experimental evidence that suppressive ODNs inhibit the development of atherosclerosis through inhibition of the STAT1/4 and T-bet pathways, which further modulate the Th1/Th2 balance in vivo.     

Impaired IL-4 and c-Maf expression and enhanced Th1-cell development in Vav1-deficient mice

Although c-Maf is crucial for Th2 differentiation and production of interleukin 4 (IL-4), its regulation is poorly understood. We report that Vav1-/- CD4+ T cells display deficient T-cell receptor (TCR)/CD28-induced IL-4 and c-Maf expression and, conversely, enhanced interferon gamma (IFN-gamma) production and T-bet expression (even when cultured under Th2-polarizing conditions), but intact expression of other Th2 cytokines and GATA-3. Up-regulation of c-Maf was dependent on Ca2+/nuclear factor of activated T cell (NFAT) and, together with IL-4 production, could be rescued in Vav1-/- T cells by Ca2+ ionophore. Deficient IL-4 production was restored by retrovirus-mediated Vav1 expression, but only partially by retroviral c-Maf expression. Similar IL-4 --> IFN-gamma skewing was observed in intact, antigen-primed Vav1-/- mice. Thus, Vav1 is selectively required for IL-4 and c-Maf expression, a requirement reflecting, at least in part, the dependence of c-Maf expression on Ca2+/NFAT signaling.     

支气管哮喘患者Th2细胞过度分化与转录因子T-bet和GATA-3的调控作用

目的　探讨支气管哮喘 (简称哮喘 )患者是否存在Th2细胞过度分化以及转录因子T bet和GATA 3的调控作用。方法　将 32例哮喘患者 (A组 )和 2 0名健康对照者 (B组 )纳入研究。A组中儿童型 (A1组 )和成人型 (A2 组 )分别为 12、2 0例 ,变应原皮试阳性 (AⅠ 组 )和阴性 (AⅡ 组 )者分别为 18、14例。采用酶联免疫吸附测定 (ELISA)法检测哮喘患者外周血淋巴细胞培养上清液中白细胞介素 4 (IL 4 )和γ干扰素 (INF γ)浓度 ,用直接免疫荧光标记法测定淋巴细胞中CCR3 和CCR5 阳性细胞率 ,用逆转录 聚合酶链反应 (RT PCR)和流式细胞术 (FCM)测定淋巴细胞中T bet、GATA 3mRNA表达水平。结果　A组和B组患者淋巴细胞培养上清液中IL 4、INF γ浓度分别为 (118± 2 5 ) μg/L、(75± 12 ) μg/L、(6 5 1± 85 ) μg/L、(1179± 332 ) μg/L ,A、B两组比较差异均有显著性 (P均 <0 0 0 1) ;A1组和A2 组淋巴细胞培养上清液中IL 4浓度分别为 (12 1± 2 5 ) μg/L、(118± 2 5 ) μg/L ;INF γ浓度分别为 (6 39±132 ) μg/L、(6 6 1± 84 ) μg/L ,A1和A2 组分别与B组比较差异均有显著性 (P均 <0 0 1) ,但A1组与A2 组间比较差异无显著性 (P >0 0 5 )。AⅠ 组与AⅡ 组IL 4浓度分别为 (12 6± 2 3) μg/L、(10 7± 2 6     

T-bet、GATA-3在慢性特发性血小板减少性紫癜患者外周血中的表达及意义

目的进一步探讨慢性特发性血小板减少性紫癜（CITP）的发病机制。方法选择25例CITP患者（CITP组）及25例健康体检者（正常对照组）,采用ELISA法检测外周血Th细胞因子IFN-γ、IL-10表达;采用RT—PCR检测外周血淋巴细胞中转录因子T-bet、GATA-3mRNA表达。结果与正常对照组相比,CITP组IFN—γ表达显著升高、IL-10表达显著降低（P〈0．01）,T—betmRNA表达明显升高、GATA-3mRNA表达明显下降（P〈0．05）。结论T-bet、GATA-3表达异常在CITP发生、发展过程中发挥重要作用,可能机制为增强TM细胞功能、抑制Th2细胞功能。     

支气管哮喘患者CC16与气道炎症及Th1/Th2细胞因子的关系

目的探讨支气管哮喘（哮喘）患者CC16与气道炎症及Th1/Th2细胞因子的关系。方法采取病例对照研究,收集25例哮喘急性发作期患者和33例健康对照组。采外周静脉血分离血浆,提取外周血单个核细胞（PBMC）,用ELISA测定血浆中CC16、干扰素-γ（IFN-γ）、白介素-4（IL-4）的水平;用RT-PCR法检测PBMC转录因子T-bet和Gata-3mRNA表达水平。结果 1.哮喘组CC16及IFN-γ分别为（21.96±7.31）ng/ml,（118.73±22.59）pg/ml,明显低于对照组[分别为（64.88±25.27）ng/ml和（145.53±29.50）pg/ml,（均P〈0.01）],哮喘组IL-4（425.22±4.37）pg/ml高于对照组（69.72±10.15）pg/ml,（P〈0.01）。2.哮喘组T-betmRNA、T-bet/GATA-3表达水平（0.12±0.01,0.25±0.04）显著低于对照组（0.48±0.12,1.894±0.65）（均P〈0.01）,GATA-3mRNA表达（0.45±0.05）较对照组明显升高（0.30±0.08）（P〈0.01）。3.CC16与T-betmRNA表达水平、T-bet/GATA-3呈正相关（r分别为0.792,0.761,均P〈0.01）;与GATA-3mRNA无明显相关性（r=-0.146,P=0.551）。结论 CC16参与哮喘气道炎症反应,并以Th1/Th2细胞因子失衡为特点。     

免疫性血小板减少症患者外周血白细胞介素-18及其受体α链的表达

目的 通过检测新诊断的免疫性血小板减少症(ITP)患者用药前外周血IL-18和CD_3~+细胞表面IL-18受体α链(IL-18Rα)的表达,探讨IL-18和IL-18Rα在ITP发病中的作用机制.方法 以我院门诊及住院治疗的18例新诊断的ITP为研究对象,应用ELISA检测血浆中IL-18的含量,采用流式细胞术分析CD_3~+细胞和总淋巴细胞表面IL-18Rα的表达,应用RT-PCR检测外周血单个核细胞(PBMCs)中IL-18 mRNA、转录因子T-bet mRNA和GATA-3 mRNA的表达.选择15例与试验组匹配的健康志愿者作为正常对照组.结果 新诊断的ITP患者血浆中IL-18的含量为(468.57±141.62)pg/ml,显著高于对照组(P<0.05);CD_3~+细胞表面及淋巴细胞表面IL-18Rα的表达分别为(8.50±3.16)%和(9.16±2.98)%,两者均显著高于对照组(P<0.05);ITP患者PBMCs中IL-18 mRNA、T-bet mRNA和GATA-3 mRNA的表达分别为0.12±0.02、0.07±0.02和0.0039±0.0014,IL-18mRNA和T-bet mRNA显著高于对照组(P<0.05),而GATA-3 mRNA显著低于对照组(P<0.05),T-bet/GATA-3比例显著增高.结论 IL-18与ITP的发生发展有关,本研究从蛋白和基因水平说明IL-18和IL-18Rα可能上调ITP患者Th1类细胞的表达,通过调整T-bet/GATA-3的比例,恢复Th1/Th2的平衡,有望为ITP的治疗提供一个新的治疗策略.