过继转输扩张型心肌病小鼠淋巴细胞诱导心肌损害

目的探讨扩张型心肌病(DCM)小鼠的自身免疫学机制。方法用人线粒体ADP/ATP载体肽免疫小鼠建立扩张型心肌病模型(DCM组),分别将其以伴刀豆凝集素A(ConA)刺激和未刺激的脾淋巴细胞过继转移给同源近交系小鼠(DCM-T-A组和DCM-T组),以用不含肽的对照液体免疫的小鼠为对照组;观察各组小鼠心肌组织病理学变化,以酶联免疫吸附法检测其血清抗ADP/ATP载体自身抗体水平、实时荧光定量聚合酶链反应检测其心肌细胞因子基因表达。结果在免疫或过继后的第1和第6个月末,DCM组和以ConA刺激的DCM-T-A组小鼠血清抗ADP/ATP载体自身抗体均为阳性;心脏在前期分别出现心肌炎性细胞浸润和轻度心肌纤维化,在后期均出现心脏增大、心肌细胞溶解断裂和弥漫性纤维化;心肌中细胞因子表达均增高,其中干扰素(IFN)-γ(DCM组30d:1486.0±243.0,180d:192.0±38.0;DCM-T-A组30d:357.0±38.6,180d:186.0±19.27)和白细胞介素(IL)-2(DCM组30d:197.0±11.0,180d:103.0±19.0;DCIN-T-A组30d:166.0±13.8,180d:96.0±4.47)表达增高在前期明显,IL-4、IL-6和肿瘤坏死因子(TNF)-α表达增高在后期更为显著。未用ConA刺激的DCM-T组与对照组均无上述改变。结论过继转输DCM小鼠脾淋巴细胞可诱导同源近交系小鼠产生抗ADP/ATP载体自身抗体和T细胞介导的免疫应答,发生过继免疫性心肌损害,且此损害与供体小鼠体内改变相似。     

过继转输抗L3T4单抗治疗的心肌病小鼠脾细胞对心肌抗体的影响

目的 通过对心肌自身抗体的分析探讨Th细胞在自身免疫性心肌病发病机制中的作用.方法 用含有人线粒体ADP/ATP载体肽的免疫液多次免疫Balb/c小鼠得到心肌病组(n=6),单抗组(n=6)小鼠在给予ADP/ATP肽免疫的第0、1和2天,同时接受尾静脉注射400 μg抗L3T4单抗;第180天时,将单抗组小鼠的脾细胞取出转输到正常小鼠体内(转输组,n=6),此小鼠亦同时给予多肽免疫,方法同心肌病组.对照组(n=6),以不含多肽的盐水免疫小鼠,时间和剂量同心肌病组.以ELISA法检测各组小鼠血清抗ADP/ATP载体自身抗体水平及IgG亚型的变化;以实时荧光定量PCR法检测各组小鼠T细胞中的细胞因子mRNA的表达.结果 转输组小鼠血清自身抗体在各时间点均为阴性,类似于单抗组和对照组,而心肌病组抗体水平在各时间点均显著高于对照组(P<0.01);转输组IgG1亚型的生成受到明显抑制;细胞因子(IFN-γ、IL-2、IL-4和IL-6)表达在单抗组和转输组均受到显著抑制.结论 Th细胞介导的免疫应答在自身免疫性心肌病的发病机制中起重要作用.     

扩张型心肌病患者血清中抗ADP/ATP载体抗体的临床意义

扩张型心肌病(DCM)病因不明,近年来大量资料证实,DCM与病毒核糖感染及自身免疫有关.DCM患者血清中存在肠病毒核糖核酸(RNA);其血清中还存在器官特异性抗心肌抗体,并介导心肌自身免疫损害.本文应用免疫转印技术,探讨DCM患者血清中抗ADP/ATP载体抗体的临床意义.     

抗L3T4单克隆抗体对小鼠自身免疫性心肌病免疫干预的研究

目的探讨抗L3T4单克隆抗体(单抗)对自身免疫性心肌病的免疫治疗作用。方法以线粒体ADP/ATP载体肽免疫小鼠建立自身免疫性心肌病模型(心肌病组,6只);早期治疗组小鼠(6只)在给予肽免疫的前1d、当天和后1d,同时接受尾静脉注射抗L3T4单抗;中期治疗组小鼠(6只)在给予肽免疫的第3个月接受抗L3T4单抗的治疗;以不含肽的空白免疫液免疫小鼠为对照组(6只)。采用酶联免疫吸附法(ELISA)检测小鼠血清抗线粒体ADP/ATP载体抗体动态变化,以流式细胞术检测小鼠T细胞中细胞因子干扰素(IFN)-γ和白细胞介素4(IL-4)的表达,观察小鼠心肌组织在光镜和电镜下的病理改变,计算组织中的胶原纤维容积分数。试验期半年。结果早期治疗组与对照组小鼠抗线粒体ADP/ATP载体抗体检测均为阴性,心肌组织无病理变化;而中期治疗组与心肌病组抗体均为阳性,心脏出现类扩张型心肌病样改变,但前者抗体水平有短暂性下降,且心肌病变较后者稍轻。早期治疗组T细胞中IFN-γ/IL-4百分含量(6·56%±0·21%/2·54%±0·20%)与对照组(5·87%±0·19%/2·16%±0·10%)相比,差异均无统计学意义;而中期治疗组(8·91%±0·33%/5·15%±0·13%)与心肌病组(7·85%±1·42%/9·45%±1·70%)均高于对照组,二者与前两组相比,差异均有统计学意义。结论抗L3T4单抗对线粒体ADP/ATP载体肽诱导的小鼠自身免疫性心肌病有治疗作用,且早期疗效更为明显。     

扩张型肌病患者血清中抗ADP／ATP载体抗体的临床意义

扩张型，心肌病(DCM)病因不明，近年来大量资料证实，DCM与病毒核糖感染及自身免疫有关。DCM患者血清中存在肠病毒糖碳酸（RNA)；其血清中还存在器官特异性抗心肌抗体，并介导心肌自身免疫损害。本文应用免疫转印技术，探讨DCM患者血清中抗ADP/ATP载体抗体的临床意义。     

地尔硫()治疗扩张型心肌病24例疗效观察

扩张型心肌病(DCM)的病因及发病机理不明.近年来的研究表明,DCM患者体内存在一种抗心肌线粒体ADP/ATP载体抗体,这种自身抗体可以引起实验心肌细胞毒性损伤[1,2],这种细胞损伤具有钙依赖性,钙拮抗剂可以降低这种毒性反应[3].本文旨在观察钙拮抗剂-地尔硫对扩张型心肌病的心肌保护作用.     

扩张型心肌病分子免疫治疗靶点的研究进展

扩张型心肌病（DCM）是导致心力衰竭的主要心肌疾病之一,其发病率有逐年增长和低龄化趋势,针对心力衰竭治疗是该病治疗的基本方法。随着临床和实验研究的深入,已发现病毒持续感染、免疫应答异常和遗传因素与扩张型心肌病的发生密切相关,尤其近十余年来一系列抗心肌自身抗体（包括抗线粒体ADP／ATP载体抗体、抗β1-肾上腺素能受体抗体、抗肌球蛋白抗体、抗M2-胆碱能受体抗体等）在该病患者体内的发现及其作用机制的阐明,充分证明自身免疫机制在其发病中起重要作用。     

Analysis of Specific Th1/Th2 Helper Cell Responses and IgG Subtype Antibodies in Anti-CD4 Monoclonal Antibody Treated Mice with Autoimmune Cardiomyopathy

The cytokine repertoire of ADP/ATP carrier-specific humoral immune responses and the cytokine-dependent anti-ADP/ATP carrier antibody IgG subclasses were examined in a cohort of ADP/ATP carrier-immunized BALB/c mice treated with anti-CD4 monoclonal antibody. Eighteen male BALB/c mice （6–8 weeks old） were randomized into 3 groups： dilated cardiomyopathy （DCM） group, DCM-tolerance （Tol） group and control group. The mice in DCM group were immunized with the peptides derived from human ADP/ATP carrier protein for 6 months and mice in the control group were sham-immunized, while the mice in DCM-Tol group were immunized with ADP/ATP carrier protein and anti-CD4 McAb simultaneously. Serum autoantibody against ADP/ATP carrier and IgG subclasses were measured by ELISA, intracellular cytokines IFN-γ and IL-4 of Th cells were moni- tored with flow cytometry, and splenic T cell cytokines IFN-γ, IL-2, IL-4 and IL-6 were detected by using real-time fluorescent quantitative PCR. The results showed that the autoantibody against ADP/ATP carrier was found in all mice in DCM group, and the antibody level, serum IgG1 and IgG2a subclasses, cytokines in T cells and Th cells were all elevated in DCM group, as compared with those in control group （P〈0.01）. On the other hand, in DCM-Tol group, the autoantibody level and contents of all the cytokines were significantly different from those in DCM group （P〈0.01）, and were close to those in control group. And the levels of IgG1, IgG2a, IgG2b and IgG3 were influenced, to varying degrees, by anti-CD4 McAb as compared with those in DCM group. All these four types of IgG subclasses were substantially decreased in DCM-Tol group as compared with DCM group. It is concluded that the treatment with anti-CD4 McAb could prevent the activation of T cells, reverse the abnormal secretion of cytokines and the imbalance between Th1/Th2 cell subsets and abnormal production of autoantibody against ADP/ATP carrier, and eventually avoid myocardial injuries.     

不同品系小鼠感染华支睾吸虫后IgG及亚类的动态观察

目的观察不同品系小鼠感染华支睾吸虫后，血清中特异性抗体IgG、IsGI／IgG2a的动态变化。方法NaOH消化法粪检虫卵，ELISA法测定不同品系小鼠感染后4、8和12w血清中抗体IsG、IsGI／IgG2a的动态变化。结果三个品系小鼠血清IsG均于感染后第4～8w升高，约在第10w达到高峰，此后开始下降并维持至观察结束的第12w；IgGI／IgG2a比值感染后逐渐增高，并持续至第12w。结论华支睾吸虫感染的免疫应答发生了由Th1免疫应答向Th2免疫应答转变的趋势。     

合成肽抗原检测ADP/ATP载体抗体及其在病毒性心脏病中的应用

目的 探讨心肌细胞线粒体内膜ADP/ATP载体（ANT）抗体在病毒性心脏病（VHD）诊断中的价值。方法 通过计算机预测ANT抗体决定簇的氨基酸序列,并人工合成多肽片段,以此片段作为抗原,采用酶联免疫吸附法检测急性病毒性心肌炎（AVMC）21例、慢性病毒性心肌炎（CVMC）54例、扩张型心肌病（DCM）41例及健康献血员45例。结果 CVMC患者中ANT抗体阳性率为57.4%（31/54）,DCM患者阳性率为56.1%(23/41),AVMC患者阳性率为9.5%(2/21),而健康人阳性率为4.4%(2/45)。结论 合成肽检测ANT简便、快捷,可作为VHD诊断的新方法,有助于VMC和DCM的早期诊断。     

Possible Association of HLA—DRB1 Gene with the Autoantibody against Myocardial Mitochondria ADP／ATP Carrier in Dilated Cardiomyopathy

Dilated cardiomyopathy( DCM) is a heartdis-ease of unknown etiology.It's clinical feature isthat the ventricle isdilated and the systolic ventric-ular function is impaired.In recent years,manystudies have indicated that the etiology of DCM isassociated with genetic factors,virus infection andautoimmunity.Many familial DCM have beenfound,in addition to the well- recognized familialoccurrence of DCM[1] ,A genetic component hasbeen recently identified in up to2 0 % of presumedsporadic cardio…     

抗L3T4单抗早期干预对心肌病小鼠细胞因子的影响

目的研究早期应用抗L3T4单抗对心肌病小鼠血清和心肌组织中细胞因子的影响，探讨抗L3T4单抗治疗自身免疫性心肌病的机制。方法用含有人线粒体ADP／ATP载体肽的免疫液免疫近交系Balb／C小鼠建立类扩张型心肌病模型（心肌病组）；以不合肽免疫液免疫小鼠作为对照组；在同时用含有ADP／ATP载体肽的免疫液免疫小鼠的前1d连续3d以400μg抗L3T4单抗免疫小鼠获得单抗组。采用流式细胞术检测小鼠脾脏中CD44T细胞内IFN-γ／IL-4的表达；以ELISA法检测其血清中IFN-γ、IL-2、IL-4、IL-6和TNF-α水平；实时荧光定量PCR法检测其心肌细胞因子基因表达。结果心肌病组小鼠血清IFN-γ和IL-4水平明显高于对照组，而单抗早期干预使其生成受到抑制，此结果与流式细胞仪检测到的T细胞内IFN-γ和IL-4水平相-致；血清中IL-2含量在3组小鼠差异均无显著性；TNF-α则在单抗组显著高于对照组和心肌病组。从心肌组织中细胞因子表达情况来看，心肌病组各种细胞因子合成均增多，而早期应用单抗能够抑制其基因表达。结论抗L3T4单抗能够阻断绝大部分细胞因子的生成，从而能够治疗实验鼠自身免疫性心肌病。     

尼可地尔对气道平滑肌细胞增殖表达及Th1/Th2免疫失衡的影响

目的研究气道平滑肌细胞（ASMCs）增殖过程中TGF-β1分泌释放水平与Th1/Th2细胞分化的特异性细胞因子的关系,并探讨KATP通道开放剂对哮喘气道重塑的作用机制。方法体外构建增殖型ASMCs与淋巴细胞共培养的细胞模型,KATP通道开放剂尼可地尔干预,并设立对照;ELISA法检测上清液中TGF-β1的释放水平;流式细胞表面抗原法检测共培养体系中的Th1/Th2的表达差异。结果 TGF-β1的表达与ASMCs增殖呈正相关,而尼可地尔可下调TGF-β1的表达（P〈0.01）;增殖型AMSCs促进CD4＋T细胞由Th1向Th2分化,Th2优势应答,IL-4表达升高,Th1型细胞因子IFN-γ减少,Th1/Th2平衡失调;尼可地尔可作用Th1细胞百分率升高,而Th2细胞百分率降低（P〈0.05）。结论尼可地尔通过抑制ASMCs增殖而调控TGF-β1分泌表达,并可以逆转Th1/Th2免疫失衡。     

A therapeutic anti-CD4 monoclonal antibody inhibits T cell receptor signal transduction in mouse autoimmune cardiomyopathy

Background T cell immune abnormalities in patients with dilated cardiomyopathy (DCM) has been intensively studied over the past 10 years. Our previous study has suggested that immunization of mice with the peptides derived from human adenine nucleotide translocator (ANT) result in the production of autoantibodies against the ANT and histopathological changes similar to those in human DCM. The ANT peptides can induce autoimmune cardiomyopathy like DCM in Balb/c mice. In this study we aimed to focus on the molecular mechanism of T cells in the autoimmune cardiomyopathy mouse model by detecting the expression of the two T cell signaling molecules.Methods The ANT peptides were used to cause autoimmune cardiomyopathy in Balb/c mice. Anti-L3T4 or rat anti-mouse IgG was administered to the mice (n=6 in each group) simultaneously immunized with ANT. ELISA analysis was used to detect autoantibodies against the ANT peptides and the percentages of interferon-γ and interleukin-4 producing cells among splenic CD4(+) lymphocytes was determined by using flow cytometry analysis. The expression of CD45 in spleen T cells was determined by immunohistochemistry and the mRNAs of T cell signaling molecules were detected by real-time PCR.Results Treatment of ANT immunized Balb/c mice with anti-CD4 mAb caused a reduction in the gene expression of P56lck and Zap-70 and a lower level of CD45 expression by spleen T cells. Also, a reverse of the Th1/Th2 ratio that results in the reduced production of antibodies against ANT was found in the anti-CD4 monoclonal antibodies (mAb) group. Whereas irrelevant antibody (rat anti-mouse IgG) did not suppress T cell signaling molecules nor inhibit CD45 expression, and control-antibody mice did not show any significant differences compared with the DCM group.Conclusion The results show that anti-CD4 mAb is a powerful inhibitor of the early initiating events of T cell receptor (TCR) signal transduction in mouse autoimmune dilated cardiomyopathy.     

溃疡性结肠炎患者Th17和Th1／Th2细胞亚群的研究

目的检测溃疡性结肠炎（Ulcerativecolitis,UC）患者外周血中辅助性T细胞亚群（Th1、Th2和Th17）数量和比例的变化,探讨其在UC发病机制中的作用。方法选择40例UC患者,分离外周血单核细胞（PBMc）,采用流式细胞术（FCM）检测Th1、Th2、Th17细胞的数量及Th1／Th2比值,用ELISA测定培养上清中IFN-γIL-4、IL-17水平,与同期选择的健康对照组比较并与患儿的病情进行相关性分析。结果UC患者PBMC中IFN-γ＋Th1细胞的数量、IL--γ7＋Th17细胞的数量均高于健康对照组[（27．0±2．9）％,（23．2±1．7）％,P〈O．05;（26．61±2．6）,（11．4±1．2）,P〈0．011;Th1／Th2比值高于健康对照组[（12．8±1．6）,（8．60±1．9）,P〈0．01],Th17细胞的数量与疾病程度呈正相关（P〈0．05,r=0．42）。结论IL-17＋Th17细胞与Th1／Th2比值变化可能参与UC患者的免疫学发病机制。     

OSAHS患儿血清Th1／Th2细胞因子检测及临床意义

目的 探讨阻塞性睡眠呼吸暂停低通气综合征（OSAHS）患儿血清Th1/Th2免疫平衡相关细胞因子的检测及临床意义.方法 收集61例OSAHS患儿内镜下腺样体阻塞程度数据,并测定患儿血清中Th1细胞因子-肿瘤坏死因子β（TNF-β）和干扰素-γ（IFN-γ）,Th2细胞因子IL-10、IL-13和IL-12的水平.同时收集26名正常儿童样本为对照组,进行相同血清细胞因子检测.结果 OSAHS患儿血清IFN-γ的水平要明显低于正常对照组儿童,差异有统计学意义（P〈0.01）;但OSAHS 患儿其他血清Th1/Th2免疫平衡相关细胞因子（TNF-β、IL-10、IL-13和IL-12）与对照组之间差异无统计学意义（P〉0.05）.OSAHS患儿IFN-γ水平与其腺样体肥大阻塞后鼻孔程度之间并无相关性.结论 Th1和Th2免疫反应间存在制衡机制.儿童OSAHS病例中,血清IFN-γ的水平下降表明,由此导致Th1细胞介导的保护性细胞免疫反应下降,Th1/Th2失衡,从而使儿童腺样体扁桃体过度肥大而发病.     

OSAHS患儿血清Th1/Th2细胞因子检测及临床意义

目的 探讨阻塞性睡眠呼吸暂停低通气综合征（OSAHS）患儿血清Th1/Th2免疫平衡相关细胞因子的检测及临床意义。方法 收集61例OSAHS患儿内镜下腺样体阻塞程度数据,并测定患儿血清中Th1细胞因子—肿瘤坏死因子β（TNF-β）和干扰素-γ（IFN-γ）,Th2细胞因子IL-10、IL-13和IL-12的水平。同时收集26名正常儿童样本为对照组,进行相同血清细胞因子检测。结果 OSAHS患儿血清IFN-γ的水平要明显低于正常对照组儿童,差异有统计学意义（P<0.01）;但OSAHS 患儿其他血清Th1/Th2免疫平衡相关细胞因子（TNF-β、IL-10、IL-13和IL-12）与对照组之间差异无统计学意义（P>0.05）。OSAHS患儿IFN-γ水平与其腺样体肥大阻塞后鼻孔程度之间并无相关性。结论 Th1和Th2免疫反应间存在制衡机制。儿童OSAHS病例中,血清IFN-γ的水平下降表明,由此导致Th1细胞介导的保护性细胞免疫反应下降,Th1/Th2失衡,从而使儿童腺样体扁桃体过度肥大而发病。