肺炎衣原体(Cpn)诱发肺癌大鼠模型

 目的 初步探索肺炎衣原体(Chlamydia pneumoniae,Cpn)诱发大鼠肺癌模型的建立。方法 应用多次气管内注入Cpn菌液(TW 183)的方法试制肺癌大鼠模型,微量免疫荧光(microimmunofluorescence,MIF)法检测大鼠血清中Cpn特异性抗体,PCR检测肺癌组织中Cpn DNA,酶联免疫法检测肺癌组织中Cpn特异性抗原。结果 单独Cpn感染组Wistar鼠的Cpn感染率为72.9%(35/48),Cpn感染联合苯并芘(benzopyrene,Bp)组的Cpn感染率为76.7%(33/43),两组的肺癌发生率分别为25.0%(12/48)和44.2%(19/43),与正常对照组相比,P值分别为0.001和0.000。结论 Cpn感染诱发大鼠肺癌模型的建立是成功的,有助于肺癌的防治及其发病机制研究。     

Chaperonin 10 in the rat oocytes and early embryos: its expression and activity for early pregnancy factor

PROBLEM: The aim of this study was to investigate the expression of chaperonin (cpn) 10 and cpn 60 mRNA in oocytes or embryos, and to further explore the possibility that early pregnancy factor (EPF) is identical with cpn 10. METHOD OF STUDY: The expressions of cpn 10 and cpn 60 mRNA in oocytes and embryos at the different stages (1-cell, 2-cell, 8-cell, and morula) were examined by polymerase chain reaction techniques. The EPF activity of native rat cpn 10 isolated from rat livers was evaluated by the rosette inhibition test. RESULTS: Similar levels of mRNA of cpn 10 and cpn 60 were detected in oocytes and embryos at every stage. There were no detectable EPF activities in the native cpn 10. Immunoprecipitation using polyclonal antibodies against cpn 10 did not affect the activity of EPF in the pregnant rat serum. CONCLUSION: Our results do not support the hypothesis that cpn 10 is identical with EPF.     

噻托溴胺联合阿奇霉素治疗慢性阻塞性肺疾病

目的：观察噻托溴胺联合阿奇霉素治疗慢性阻塞性肺疾病（COPD）的临床疗效和肺功能变化。方法：选择COPD急性加重期住院患者50例（第1组）,COPD稳定期患者42例（第2组）,对照组为同期参加呼吸疾病体检的老年人65例。分离外周血单核细胞（PBMC）,用直接免疫荧光（DIF）法检测PBMC中的肺炎衣原体（Cpn）特异性抗原（Ag）,同时用间接微量免疫荧光（MIF）法检测Cpn抗体（IgA,IgG和IgM）。胶体金法快速定量测定C-反应蛋白（CRP）。结果：第1组Cpn的感染率为66%（33/50）,第2组为66.7%（28/42）,对照组为21.5%（14/65）。Cpn-Ag和Cpn-IgM抗体的检出率第1组显著高于对照组（P〈0.001）,Cpn-IgA和IgG抗体的检出率第1组和第2组均显著高于对照组（P〈0.01）。治疗后第1组和第2组患者的临床症状均有显著改善：Borg记分下降和FEV1/FVC（%）显著增加（P〈0.01）;Cpn-IgM阳性率显著下降（P〈0.001）;第1组和第2组CRP水平下降均有显著差异（P〈0.01）。结论：COPD患者有较高的Cpn感染率,噻托溴胺联合阿奇霉素治疗COPD彼此有正协同效应。     

肺炎嗜衣原体Cpn 0147重组蛋白的免疫学活性研究

目的 研究Cpn 0147重组蛋白的免疫学活性及其应用于肺炎嗜衣原体(Cpn) 感染临床诊断中的价值.方法 采用GST琼脂糖凝胶纯化目的 蛋白,将Cpn 0147重组蛋白免疫新西兰兔,分别应用Western blot、ELISA法检测其免疫反应性及免疫原性,同时通过检测临床标本以评估其诊断价值.结果 成功表达并纯化了相对分子质量约41×103的重组蛋白GST-Cpn 0147;Western blot和ELISA结果显示,该重组蛋白具有良好的免疫反应性,动物实验结果表明该重组蛋白具有较好的免疫原性,ELISA结果显示该重组蛋白免疫新西兰兔血清抗体效价为1∶12 800.结论 Cpn 0147重组蛋白具有较好免疫学活性和特异性,可为Cpn感染的确诊、预防研究奠定基础.     

Oral dosing with multi-antigenic construct induces atheroprotective immune tolerance to individual peptides in mice

Inflammatory immune response to self-antigens plays an important role in the development of atherosclerosis. Restoring immune tolerance to self-proteins reduces the pro-inflammatory response. We previously showed that oral tolerance to a combination of two peptides is atheroprotective. In the present study we expressed epitopes from apolipoprotein B 100 (ApoB), human heat shock protein (HSP60) and Chlamydia pneumonia outer membrane protein (Cpn) in a single protein scaffold and used this multi-antigenic construct to induce tolerance to individual peptides by oral route in ApoB^tm2Sgy/Ldlr^tm1Her/J mice. Antigen specific tolerance to individual peptides was observed in treated animals as seen by an increase in regulatory T cells. Tolerance to the peptides resulted in a 46.5% (p = 0.002) reduction in the development of atherosclerosis compared with control. Atheroprotection was associated with a significant (p < 0.05) decrease in plaque inflammation and an increase in the expression of immune regulatory markers in the aorta. CD11c⁺ cells coexpressing CD11b and CD103 increased in lymphoid organs and were found to activate regulatory T cells and reduce effector T-cell response. Adoptive transfer of CD11c⁺ cells was atheroprotective. Our results suggest that atheroprotection by oral tolerance to a multi-antigenic construct is mediated by antigen specific regulatory T cells and CD11c⁺ cells with immune regulatory properties.     

慢性肺炎衣原体感染是慢性阻塞性肺病（COPD）的危险因子

目的研究慢性Cpn感染和慢性阻塞性肺病（COPD）之间可能的相关性。方法观察组为年龄在58岁以上的轻度至重度的COPD患者165例和年龄、性别匹配的80例对照组,并测定其FEVl、FVC和圣乔治呼吸问卷（SGRQ）计分。用直接免疫荧光（DIF）法检测外周血单核细胞（PBMC）中的肺炎衣原体特异性抗原（Cpn—Ag）,同时用间接微量免疫荧光（MIF）法检测Cpn抗体（IgA,IgG和IgM）。结果观察组Cpn的感染率为63．0％（104／165）,对照组15．0％（12／80）。代表急性感染的Cpn—Ag和Cpn-IgM抗体的检出率观察组亦显著高于对照组（P〈O．001）,代表慢性感染的Cpn—IgA和IgG抗体的检出率观察组也显著高于对照组（P％0．001）。阿奇霉素治疗后观察组病人的临床症状均有显著改善：SGRQ记分和FEV。／FVC（％）显著增加,与此同时观察组只有Cpn—IgM滴度显著下降（P〈O．001）。慢性Cpn感染与吸烟及较高的年龄有关但与性别无关。结论慢性Cpn感染可能是COPD发展的独立危险因素。     

Cpn0585蛋白的免疫活性及血清学诊断初步评价

目的克隆肺炎嗜衣原体包涵体膜蛋白Cpn0585基因,初步探讨其在血清学诊断中的应用价值。方法构建pGEX-6p-2／Cpn0585重组质粒,诱导表达并纯化重组蛋白,免疫BALB／c小鼠,间接酶联免疫附试验（ELISA）检测小鼠血清抗体滴度,以重组蛋白作为ELBA包被抗原检测血清中CpnIgG抗体和检测36份Cpn-／Ct＋IsG阳性参考血清。结果高效表达和纯化出一相对分子质量约为95kDa的重组蛋白,蛋白质印迹法证明其能与人抗CpnIgG抗体发生反应;在被免疫的BALB／c小鼠体内,特异性IgG抗体的滴度为1：12800;检测104例临床血清标本中的IgG抗体,与以色列SavyonDiagnostics公司SeroCPTMIgGELISA诊断试剂盒的检测结果进行比较,符合率为98．3％。结论表达的Cpn0585重组蛋白具有良好的免疫活性,在Cpn的血清学诊断中具有较高的应用价值。     

Diagnostic value of an ELISA using a recombinant 54-kDa species-specific protein from Chlamydia pneumoniae

The aim of this study was to evaluate a 54-kDa recombinant protein encoded by the CPn0980 gene for use in a Chlamydia pneumoniae-specific ELISA. The ability of this affinity-purified protein to detect C. pneumoniae-specific antibodies was evaluated with a panel of 105 serum samples from 62 patients with community-acquired pneumonia. The results of this assay were compared with those obtained with a direct PCR-based detection assay and an outer-membrane complex-based immunoassay. The 54-kDa protein induced specific antibodies following infection of humans, and the recombinant 54-kDa ELISA detected anti-C. pneumoniae IgG and/or IgM antibodies with a sensitivity of 66.7% and a specificity of 79.2% compared with the direct PCR-based detection assay.     

Cryptosporidium parvum Cpn60 targets a relict organelle

Chaperonin 60 (Cpn60) is a well-established marker protein for eukaryotic mitochondria and plastids. In order to determine whether the small double-membrane-bounded organelle posterior to the nucleus in the apicomplexan Cryptosporidium parvum is a mitochondrion, the Cpn60 gene of C. parvum sporozoites (CpCpn60) was analyzed and antibodies were generated for localization of the peptide. Sequence and phylogenetic analyses indicated that CpCpn60 is a mitochondrial isotype and that antibodies against it localize to the rough endoplasmic reticulum-enveloped remnant organelle of C. parvum sporozoites. These data show this organelle is of mitochondrial origin.Communicated by M. Brunner     

肺炎嗜衣原体Cpn0810重组蛋白诱导THP-1细胞产生促炎因子和凋亡的研究

目的:在E.coliBL21中表达肺炎嗜衣原体Cpn0810,并研究该蛋白能否诱导人单核细胞( THP-1)产生TNF-α和IL-6等促炎因子和细胞凋亡.方法:PCR扩增Cpn0810蛋白编码基因,构建pGEX6p-2/Cpn0810重组质粒,在E.coliBL21中诱导表达,经ToxinEraser纯化柱纯化后,用不同浓度的GST-Cpn0810作用THP-1细胞,ELISA法检测TNF-α和IL-6的水平,Hoechst33258荧光染色、AnnexinV-F1TC-PI染色法检测细胞凋亡情况.结果:构建了重组质粒pGEX6p-2/Cpn0810,并在E.coliBL21菌中高效表达.该蛋白能诱导THP-1细胞以时间、剂量依赖方式表达TNF-α和IL-6;当10mg/L GST-Cpn08 10处理THP-1细胞24h后,形态学上,细胞表现为皱缩、核碎裂、细胞起泡及凋亡小体等凋亡特征.结论:表达并纯化的Cpn0810蛋白能诱导THP-1细胞分泌TNF-α和IL-6等促炎因子和诱导其发生凋亡.