补体系统在动脉粥样硬化中的作用研究进展

炎症学说认为动脉粥样硬化(As)是由修饰的脂质颗粒与单核细胞、巨噬细胞共同参与的免疫炎症反应。补体系统作为先天性免疫的一部分,是As发生发展的重要参与者。部分补体成分在As发生发展过程中的作用有新的进展,例如,补体蛋白C1q在As中具有双重作用,即致As作用和抗As作用;补体成分C3通过与其受体结合或形成非蛋白水解中间体,参与As及晚期血栓形成;补体成分C3a与C3a受体、C5a与C5a受体1或C5a受体2结合后,通过不同途径促进核苷酸结合寡聚化结构域样受体热蛋白结构域亚家族成员3炎症小体活化、白细胞介素1β分泌,促进As发生;C5b-9补体复合物通过致动脉内皮功能障碍,从而发挥致As作用,且补体应答基因32是其关键效应因子。     

结核硬脂酸棒状杆菌肺部感染1例

<正>结核硬脂酸棒状杆菌(Corynebacterium tuberculosis, C.t.)是一种不产生芽孢且革兰染色阳性的亲脂性棒状杆菌，常为人类皮肤和黏膜表面的正常菌群，但在侵入性操作后可侵入深层组织并引起感染。C.t.感染在临床中较少见，为提高临床医生对本病的认识，通过1例培养诊断为结核硬脂酸棒状杆菌肺部感染的死亡病例报道，并结合国内外相关文献分析其临床特征及治疗方法等。病例资料患者，男，74岁，     

结核分枝杆菌ATP结合盒超家族外排泵的研究进展

近年来,结核分枝杆菌的耐药菌株日益增多,使得治疗难度也在加大.其耐药主要是通过细胞壁的阻碍作用和基因突变所导致.但是仅凭这两种因素不能够完全解释分枝杆菌的耐药机制.据研究,分枝杆菌的主动外排是另一种重要的耐药机制.主动外排泵对结核分枝杆菌的耐药方面的影响也越来越受人们的关注,而外排泵的类型有很多,其中ATP结合盒超家族是外排泵家族中的大家族,在外排作用中起到了重大影响,通过总结近年来发表的文献,现对结核分枝杆菌ATP结合盒超家族的抑制剂和作用底物的研究进行了综述.     

结核分枝杆菌药敏试验方法改进的探讨

目的 探讨改进结核分枝杆菌药敏试验方法?方法 以 B A C T E C 法7 H12 B 液体培基分离出50例临床结核分枝杆菌菌株,经不同稀释后,直接接种于改良罗氏药敏培基上,与 B A C T E C 法药敏结果对比,观察其耐药性?结果 1 .7 H12 B 液体培基中结核分枝杆菌菌液 G I 值500 ～900 时,1 ml 相当于1 mg 结核分枝杆菌湿重的活菌( 菌数106 ～7) ?2 .取7 H12 B 液体培基的菌液直接接种于罗氏药敏培养基,其耐药结果与 B A C T E C 法相比,无显著性差异( P> 0.05) ?结论 此法可弥补 B A C T E C 通常只有四种药敏的不足,与常规罗氏药敏方法比较,可提前报告13 ～15 天?     

308例结核抗体快速测定分析

用结核抗体快速测定试剂盒对 30 8例诊断为结核病和非结核病例进行测试 ,观察结核病活动期、静止期以及非结核病例中结核抗体 Ig G的水平。1　临床资料2 0 0 0年 3月至 12月确诊 199例 (6 4 .6 % )活动性结核患者 ;10 9例 (35 .4 % )非结核病和静止期结核患者 ,门诊和住院病例各占 15 4例。其中男性 16 3例 ,平均 35 .7岁 ;女性 14 5例 ,平均 34岁。男女平均 37.4岁 ,性别比为 1∶ 1.12。2　方　法采用结核纯化蛋白衍生物为抗原 ,包被聚苯乙烯反应板微孔 ,待测样品中有结核抗体 ,可与之结合 ,再与酶标抗体反应 ,经酶底物溶液作用即可呈色 ,…     

肿瘤坏死因子-α拮抗剂引发结核二例分析并文献复习

目的 提高临床医生对肿瘤坏死因子(TNF)-α州拈抗剂治疗引发结核的认识.方法 报道2例类风湿关节炎(RA)患者经TNF-α拮抗剂治疗后发生结核病的临床情况,并复习近年国内外相关文献.结果 病例1在3个月内接受4次英夫利昔3 ms/ks静脉注射治疗后出现右锁骨上淋巴结结核,手术切除及四联抗结核治疗后痊愈.病例2接受依那西普25 mg皮下注射2次/周治疗1.5个月后发生肺结核瘤,手术切除后好转.文献复习显示TNF-α拮抗剂治疗可增加结核的发病率,英夫利昔的致结核作用强于依那西普,发生结核的患者多为老年人,肺外结核及播散性结核的发生率较高.结论 TNF-α拮抗剂治疗可增加结核的发病率,治疗前结核筛查、治疗期间及治疗后监测结核的发生很有必要.     

肺泡表面相关蛋白A、D与结核分枝杆菌感染相关性的研究进展

肺泡表面活性物质是由肺泡上皮细胞及支气管上皮细胞产生和分泌的,主要分为SP-A、SP-B、SP-C、SP-D等,在肺内免疫调节及炎症反应方面有重要作用,其中最主要的是SP-A和SP-D.肺结核是机体对结核分枝杆菌侵犯所产生的一种变态和免疫反应同时存在的状态,结核分枝杆菌在侵入肺泡之前首先要经呼吸道,被肺泡巨噬细胞吞噬,而肺泡表面活性物质在结核分枝杆菌与肺泡巨噬细胞之间起着重要中介体的作用.SP-A能够识别结核分枝杆菌细胞壁上的受体或者其他糖类分子,SP-D可以与结核分枝杆菌表面的脂质阿拉伯甘露聚糖结合,它们在介导结核分枝杆菌与肺泡巨噬细胞的黏附时受到一定因素的调节及限制,最终决定结核分枝杆菌与肺泡巨噬细胞之间黏附和吞噬的程度,从而决定结核分枝杆菌进入机体后增殖或被降解的结局.     

细胞因子基因多态性与结核病易感性研究进展

结核病是发展中国家常见的疾病,病死率高.全球有近三分之一的人口感染结核分枝杆菌,但仅有10%的感染个体最终发展为临床结核,说明宿主的基因因素在结核病的发病机制中起着极其重要的作用.细胞因子是结核病免疫的重要组成部分,其水平与结核病易感性、严重程度和临床表现都有联系,因此编码细胞因子的基因在结核病易感性中有重要的作用.现就细胞因子基因多态性与结核易感性的关系作一综述.     

流式检测C1_q~+的细胞表达及在肺结核诊断中的价值

目的初步建立全血细胞中CD+16CD+68CD+14CD-3C1+q的流式检测方法,评价其在结核病诊断中的价值。方法利用流式细胞技术检测CD+16CD+68CD+14CD-3细胞表面C1+q的表达,比较其在健康照(26例),结核潜伏感染者(12例)、活动性结核患者(19例)的表达情况。结果活动性结核患者组外周血CD+14CD-3细胞显著高于健康对照(P0.05);活动性结核患者组CD+16CD+68CD+14CD-3细胞表面C1+q的表达显著高于结核潜伏感染组和健康对照组(P0.05)。结论在活动性结核病人的外周血中,C1+q在CD+16CD+68CD+14CD-3细胞表面高于潜伏感染者,为活动性肺结核患者与潜伏感染者的临床鉴别诊断技术的研究和开发,提供了一条可行的思路。     

The analysis of image characteristics of 18F-FDG PET/Cr in untypical tuberculosis

目的 探讨不典型结核病18氟-氟代脱氧葡萄糖(18F-FDG)正电子发射断层扫描(PET/CT)的显像特点.方法 分析43例通过病理或者细菌学确诊并在随访后得到证实的结核患者的18F-FDG PET/CT的病灶形态与最大标准摄取值(SUVmax).结果 43例结核患者中,静止期结核3例,活动性结核患者40例.活动期患者除有临床症状外还表现为病灶对于18F-FDG的高摄取,SUVmax为6.72±3.49.结论 活动期结核病可出现18F-FDG的高浓聚,18F-FDG PET/CT检查可有效判断病变是否活动并且确认结核累及范围.     

结核分枝杆菌肝素结合血凝素在肺结核及肺外结核中的免疫作用研究

目的探讨结核分枝杆菌肝素结合血凝黏附素（heparin-binding hemagglutinin adhesin, HBHA）在不同结核感染状态人群中免疫反应及可能的诊断价值。 方法用天然的HBHA刺激选取阴性对照、潜伏感染者、肺结核和肺外结核患者,分离外周血单个核细胞,加入共培养,收集细胞培养上清,ELISA方法检测上清中IFN-γ的浓度,运用非参数检验的方法统计分析。结果4组HBHA特异的IFN-γ分泌水平分别为49.5、781.9、341.8、214.4pg/ml,潜伏感染者组远高于其他各组,HBHA 刺激后肺结核和肺外结核之间的IFN-γ分泌水平无统计学差异（U＝1901.5,P＝0.448）。结论HBHA特异的IFN-γ分泌水平,有利于鉴别健康人群中的潜伏感染者,HBHA可能具有一定的免疫保护作用,尽管HBHA与肺外结核发病相关,其特异的细胞免疫在肺结核和肺外结核间无统计学差异。     

Differential T and B cell responses against Mycobacterium tuberculosis heparin-binding hemagglutinin adhesin in infected healthy individuals and patients with tuberculosis

Because only 10% of individuals infected with Mycobacterium tuberculosis will eventually develop disease, antigens that are recognized differently by the immune systems of infected healthy and diseased subjects may constitute potential vaccine candidates. Here, the heparin-binding hemagglutinin adhesin (HBHA) is identified as such an antigen. Lymphocytes from 60% of healthy infected individuals (n=25) produced interferon (IFN)-gamma after stimulation with HBHA, compared with only 4% of patients with active tuberculosis (n=24). In the responders, both CD4(+) and CD8(+) cells secreted HBHA-specific IFN-gamma, and the antigen was presented by both major histocompatibility complex class I and II molecules. In contrast to the reduced ability of patients with tuberculosis to produce HBHA-specific IFN-gamma, most of them (82%) produced anti-HBHA antibodies, compared with 36% of the infected healthy subjects. These observations indicate that HBHA is recognized differently by the immune systems of patients with tuberculosis and infected healthy individuals and might provide a marker for protection against tuberculosis.     

Heparin-binding hemagglutinin, from an extrapulmonary dissemination factor to a powerful diagnostic and protective antigen against tuberculosis

Interactions of Mycobacterium tuberculosis with macrophages have long been recognized to be crucial to the pathogenesis of tuberculosis. The role of non-phagocytic cells is less well known. We have discovered a M. tuberculosis surface protein that interacts specifically with non-phagocytic cells, expresses hemagglutination activity and binds to sulfated glycoconjugates. It is therefore called heparin-binding hemagglutinin (HBHA). HBHA-deficient M. tuberculosis mutant strains are significantly impaired in their ability to disseminate from the lungs to other tissues, suggesting that the interaction with non-phagocytic cells, such as pulmonary epithelial cells, may play an important role in the extrapulmonary dissemination of the tubercle bacillus, one of the key steps that may lead to latency. Latently infected human individuals mount a strong T cell response to HBHA, whereas patients with active disease do not, suggesting that HBHA is a good marker for the immunodiagnosis of latent tuberculosis, and that HBHA-specific Th1 responses may contribute to protective immunity against active tuberculosis. Strong HBHA-mediated immuno-protection was shown in mouse challenge models. HBHA is a methylated protein and its antigenicity in latently infected subjects, as well as its protective immunogenicity strongly depends on the methylation pattern of HBHA. In both mice and man, the HBHA-specific IFN-gamma was produced by both the CD4(+) and the CD8(+) T cells. Furthermore, the HBHA-specific CD8(+) T cells expressed bactericidal and cytotoxic activities to mycobacteria-infected macrophages. This latter activity is most likely perforin mediated. Together, these observations strongly support the potential of methylated HBHA as an important component in future, acellular vaccines against tuberculosis.     

分枝杆菌肝素结合血凝素研究进展

结核病是由分枝杆菌感染引起的世界范围内流行的人畜共患病。肝素结合血凝素(heparin-binding adhesin,HBHA)是分枝杆菌表达的一种蛋白质,致病性分枝杆菌的HBHA蛋白具有黏附上皮细胞、扩散感染范围等功能。其发挥黏附功能的C端结构域能够刺激机体产生有效的Th1细胞免疫反应;HBHA特异的黏膜免疫反应能够提供一定的保护效果。这提示HBHA具有制作为疫苗的潜力。而且,HBHA蛋白在活动性结核病患者和潜伏感染者中引发的免疫反应存在差异,这使其可以成为新的诊断抗原。此外,在细胞水平上,HBHA可以利用补体系统进入巨噬细胞,促使巨噬细胞向M1极化,同时能够逃避自噬并诱发线粒体损伤和内质网应激介导功能,这为结核病的致病机制提供了更多的解释。HBHA与宿主之间相互作用认识的积累,可以为HBHA的应用性研究提供依据。     

结核分枝杆菌肝素结合血凝素与纯蛋白衍化物的细胞免疫作用比较

目的 通过结核分枝杆菌肝素结合血凝素(HBHA)与纯蛋白衍化物(PPD)的免疫作用比较,探讨HBHA的免疫保护作用及诊断应用价值.方法 选取阴性对照者、潜伏感染者、肺结核患者,分离入选者的外周血单个核细胞,分为无刺激、HBHA、PPD刺激组,培养4 d后收集细胞培养上清,ELISA方法检测上清中的γ-干扰素(IFN-γ)和白介素4.结果 HBHA刺激后3组产生的IFN-γ中位数分别为49.5 ng/L、781.9 ng/L、341.8 ng/L,潜伏感染者产生的IFN-γ水平远高于阴性对照者,略高于肺结核患者.HBHA特异的IFN-γ在不同治疗时间肺结核之间有显著差异.结论 HBHA有较好的免疫原性,可以刺激潜伏感染者产生高水平的IFN-γ,潜伏感染者未发展为活动性结核病可能与HBHA的免疫保护作用有关.HBHA特异的IFN-γ释放反应有利于鉴别潜伏感染者.     

结核分枝杆菌HBHA的研究进展

HBHA是结核分枝杆菌的一种表面蛋白,它与细菌的黏附作用、诱导MФ细胞凋亡以及肺外播散型结核的发生相关,在抗肿瘤免疫中也有一定的作用;也可以作为新一代的血清学诊断标识,鉴别诊断活动性与非活动性结核病,并在开发结核新疫苗及抗结核病新药中显示出潜在的的应用价值。     

结核分枝杆菌黏附素HBHA在结核病免疫学诊断中的应用

目的 探讨结核分枝杆菌肝素结合血凝黏附素(Heparin-binding haemagglutinin adhesin,HBHA)在结核病免疫诊断方面的应用价值.方法 将在苏通培养基中培养至稳定期的卡介苗菌株(BCG)菌体超声裂解,离心后取上清并通过CL-6B层析柱,利用含0～500 mmol/L氯化钠的磷酸盐缓冲溶液(PBS)进行梯度洗脱后获得天然HBHA.以BCG基因组为模板PCR扩增结核分枝杆菌hbha基因片段,质粒pET-32a为载体,大肠杆菌作为宿主菌制备原核表达的HBHA重组蛋白.以纯化的天然HBHA蛋白和原核表达的HBHA蛋白为包被抗原,分别对肺结核组、肺外结核组、PPD阳性和PPD阴性健康对照组各47例血清通过ELISA方法检测抗HBHA的抗体水平.应用SPSS 11.5统计软件对各研究组数据进行方差齐性检验,然后进行t检验,并计算各研究组的敏感度和特异度.结果 含有375 mmol/L氯化钠的PBS洗脱液可以洗脱获得较纯的天然HBHA蛋白.利用重组HBHA所带的组氨酸标签进行特异分离、纯化.ELISA检测结果表明天然HBHA蛋白和重组HBHA蛋白、PPD阳性和PPD阴性健康对照组之间血清抗体水平差异不明显.肺结核组与肺外结核组的血清抗体ELISA检测的吸光度值在散点图中分布有明显差异,肺结核组吸光度值集中分布在0.30～0.40之间,肺外结核组吸光度值分布在0.35～0.45之间,经统计学检验结果差异具有统计学意义(t=12.224,P＜0.05).结核组(肺结核和肺外结核)与对照组(PPD阴性,PPD阳性)吸光度值分布具有显著不同,对照组全部小于0.30,结核组吸光度值则集中分布于0.30～0.45之间.经统计学检验(t=25.909,P＜0.05)血清抗体水平具有显著性差异.结论 结核分枝杆菌黏附素HBHA在结核病免疫学诊断的应用中具有较好的特异度和敏感度,有望用于结核病、尤其是肺外结核病的实验室辅助诊断.     

Effector functions of heparin-binding hemagglutinin-specific CD8+ T lymphocytes in latent human tuberculosis

BACKGROUND: Most individuals infected with Mycobacterium tuberculosis do not develop tuberculosis (TB) and can be regarded as being protected by an appropriate immune response to the infection. The characterization of the immune responses of individuals with latent TB may thus be helpful in the definition of correlates of protection and the development of new vaccine strategies. The highly protective antigen heparin-binding hemagglutinin (HBHA) induces strong interferon (IFN)- gamma responses during latent, but not active, TB. Because of the recently recognized importance of CD8(+) T lymphocytes in anti-TB immunity, we characterized the CD8(+) T lymphocyte responses to HBHA in subjects with latent TB. RESULTS: HBHA-specific CD8(+) T lymphocytes expressed memory cell markers and synthesized HBHA-specific IFN- gamma . They also restricted mycobacterial growth and expressed cytotoxicity by a granule-dependent mechanism. This activity was associated with the intracellular expression of HBHA-induced perforin. Surprisingly, the perforin-producing CD8(+) T lymphocytes were distinct from the IFN- gamma -producing CD8(+) T lymphocytes. CONCLUSION: During latent TB, the HBHA-specific CD8(+) T lymphocyte population expresses all 3 effector functions associated with CD8(+) T lymphocyte-mediated protective immune mechanisms, which supports the notion that HBHA may be protective in humans and suggests that markers of HBHA-specific CD8(+) T lymphocyte responses may be useful in the monitoring of protection.     

Regulatory T cells depress immune responses to protective antigens in active tuberculosis

RATIONALE: Tuberculosis (TB) remains a leading cause of death, and the role of T-cell responses to control Mycobacterium tuberculosis infections is well recognized. Patients with latent TB infection develop strong IFN-gamma responses to the protective antigen heparin-binding hemagglutinin (HBHA), whereas patients with active TB do not. OBJECTIVES: We investigated the mechanism of this difference and evaluated the possible involvement of regulatory T (Treg) cells and/or cytokines in the low HBHA T-cell responses of patients with active TB. METHODS: The impact of anti-transforming growth factor (TGF)-beta and anti-IL-10 antibodies and of Treg cell depletion on the HBHA-induced IFN-gamma secretion was analyzed, and the Treg cell phenotype was characterized by flow cytometry. MEASUREMENTS AND MAIN RESULTS: Although the addition of anti-TGF-beta or anti-IL-10 antibodies had no effect on the HBHA-induced IFN-gamma secretion in patients with active TB, depletion of CD4(+)CD25(high)FOXP3(+) T lymphocytes resulted in the induction by HBHA of IFN-gamma concentrations that reached levels similar to those obtained for latent TB infection. No effect was noted on the early-secreted antigen target-6 or candidin T-cell responses. CONCLUSIONS: Specific CD4(+)CD25(high)FOXP3(+) T cells depress the T-cell-mediated immune responses to the protective mycobacterial antigen HBHA during active TB in humans.     

结核分枝杆菌黏附素(HBHA)对小鼠结核病免疫治疗作用的实验研究

目的 探讨结核分枝杆菌粘附素(HBHA)对小鼠结核病实验模型的免疫治疗作用.方法 取在改良罗氏培养基上生长良好的结核分枝杆菌H37Rv两周培养物,制备成均匀菌悬液,每只小鼠尾静脉注射菌量为106 CFU.于感染2周后,分别对CPG组30 ug、HBHA组5 ug、CpG+ HBHA组(CpG 30 ug和HBHA 5 ug)及对照组腹腔注射生理盐水.于感染后4周脱颈处死小鼠,称取小鼠体重、肺脏和脾脏重量,对肺和脾组织进行菌落计数,观察肺病理改变.结果 各实验组肺组织的重量与体重和对照组相比稍有偏大,菌落计数中发现HBHA免疫治疗组明显优于空白对照组,在病理切片中对照组小鼠肺组织有大片肉芽肿形成,病理损害严重.其它各组肺部病变呈弥散分布,但病变程度明显轻于对照组,在抗酸染色中空白对照组有大量的抗酸杆菌存在,其它各组相对明显减少.结论 实验结果表明HBHA对治疗小鼠结核病变模型起一定的作用.