结核分支杆菌重组BCG疫苗研究进展

结核分支杆菌引起的结核病是一种人兽共患的慢性传染病，短程督导化疗是治疗该病的主要措施，卡介苗是预防该病的唯一疫苗，但其免疫效果极不稳定。章介绍了7种结核分支杆菌重组BCG疫苗构建及其免疫机制等方面的研究进展。     

重组BCG疫苗和结构杆菌HSP65 DNA疫苗免疫原性的 …

目的 研究重组ＢＣＧ（ｒｅｃｏｍｂｉｎａｎｔ ＢＣＧ，ｒＢＣＧ）疫苗和人结核杆菌热休克蛋白（ｈｅａｔ ｓｈｏｃｋ ｐｒｏｔｅｉｎ，ＨＳＰ）６５ ＤＮＡ疫苗对小鼠免疫应答的影响，评价两种疫苗；的免疫原性。方法 用ｒＢＣＧ疫苗和ＨＳＰ６５ ＤＮＡ疫苗免疫ＢＡＬＢ／ｃ小鼠，免疫８周时，采血、取腹腔巨噬细胞和脾脏进行０实验，以释放量检测巨噬细胞吞噬活性，以淋巴细胞刺激指数（ＳＩ）反应细胞增殖能力，以ＥＬＩ     

重组微生物—BCG疫苗及其运用前景

重组BCG疫苗是一种新型微生物疫苗。本文综述了细菌、病毒和螺旋体等微生物的重组BCG疫苗及其运用前景。     

重组BCG疫苗和结核杆菌HSP65 DNA疫苗免疫原性的比较研究

目的　研究重组BCG(recombinantBCG ,rBCG)疫苗和人结核杆菌热休克蛋白 (heatshockprotein ,HSP) 6 5DNA疫苗对小鼠免疫应答的影响 ,评价两种疫苗的免疫原性。方法　用rBCG疫苗和HSP6 5DNA疫苗免疫BALB/c小鼠 ,免疫 8周时 ,采血、取腹腔巨噬细胞和脾脏进行实验。以NO释放量检测巨噬细胞吞噬活性 ,以淋巴细胞刺激指数 (SI)反映细胞增殖能力 ,以ELISA试剂盒检测血清及脾淋巴细胞培养上清的IL 2和IFN γ的含量。结果　rBCG疫苗以 10 6CFU/鼠的剂量经皮下免疫小鼠后 ,其脾淋巴细胞的增殖能力明显高于对照组 ,BCG组和HSP6 5DNA疫苗组 (P <0 .0 5 ) ;腹腔巨噬细胞一氧化氮 (NO)的含量和血清IL 2的含量明显高于对照组 (P <0 .0 1) ;血清IFN γ的含量明显高于HSP6 5DNA疫苗组 (P <0 .0 5 ) ;其余测定参数均有升高趋势 ,但差异无显著性 (P >0 .0 5 )。而HSP6 5DNA疫苗以 5 0 μg/鼠的剂量经肌注免疫小鼠后 ,其血清IL 2的含量明显高于对照组 (P <0 .0 1) ;血清IFN γ的含量明显低于rBCG组 (P <0 .0 5 ) ;其余测定参数与其他各组相比差异均无显著性(P >0 .0 5 )。结论　本实验结果显示 ,rBCG疫苗具有良好的免疫原性 ,能明显增强BALB/c小鼠的免疫应答反应。HSP6 5DNA疫苗亦能刺激机体的免疫应答 ,但其反应强度似乎不     

结核分支杆菌Ag85A DNA疫苗免疫治疗作用的研究

目的:研究结核分支杆菌Ag85A DNA疫苗的免疫治疗作用。方法:用结核分支杆菌H37Rv腹腔内注射BALB/c小鼠 2个月后,将小鼠随机分为 5组,分别用生理盐水(A)、载体质粒(B)、卡介苗(C)、维卡菌苗(D)和 Ag85A DNA疫苗(E)免疫小鼠;用ELISA法检测血清抗体;免疫治疗2月和5月时,分别取肺、肝和脾脏观察病理改变、称取重量、作菌落计数、肺组织涂片及脾淋巴细胞转化试验。结果:DNA疫苗组抗原特异性抗体不同程度升高。免疫治疗2月时,各组鼠脾淋巴细胞转化率无显著差异;各治疗组肺组织涂片细菌数和肺菌落计数均比对照组不同程度地减少;C和B组的肺病变程度较轻。免疫治疗5月时,各治疗组鼠脾淋巴细胞转化率均显著增高;各组肺、脾细菌数无显著差别比和D组的肺未见明显病变。结论:DNA疫苗似乎具有一定的免疫治疗效果。     

支气管扩张与非结核分支杆菌

随着非结核分支杆菌(NTM)感染发病率的明显上升,尤其发达国家,其与支气管扩张的关系受到人们的密切关注.本文旨对NTM的一般特点,及与支气管扩张之间的认识进行综述,对前景进行了展望.     

支气管扩张与非结核分支杆菌

随着非结核分支杆菌(NTM)感染发病率的明显上升，尤其发达国家，其与支气管扩张的关系受到人们的密切关注．本文旨对NTM的一般特点，及与支气管扩张之间的认识进行综述，对前景进行了展望．     

异烟肼耐药结核分支杆菌katG基因分析

结核分支杆菌 (Mtuberculosis,TB)的katG基因是位于细菌染色体上的结构基因 ,全长 2 2 2 3bp ,它的编码产物是过氧化氢 过氧化物酶 ,而后者在维系异烟肼 (Isoniazid ,INH)杀菌毒性中 ,发挥重要作用。因此 ,katG基因结构的正确性是编码产生功能正常的过氧化氢 过氧化物酶的先决条件 ,也是发挥INH特有的药理作用的重要基础。通过检测katG基因的分子结构 ,尤其是检测某些关键位点的碱基突变 ,可以快速发现TB对INH耐药的变异情况 ,对及时的指导临床用药 ,具有重要的意义。作者单位 :山东省医药生物技术研究中心 (黄海南 ,韩金祥 ,高雪芹…     

重组微生物-BCG疫苗及其运用前景

重组BCG疫苗是一种新型微生物疫苗.本文综述了细菌、病毒和螺旋体等微生物的重组BCG疫苗及其运用前景.     

rpoB基因套式PCR直接测序判别结核分支杆菌利福平耐药

目的 探讨rpoB基因套式PCR直接测序判别结核分支杆菌（Mrb)利福平耐药。方法 收集肺结核患者痰标本进行rpoB基因套式PCR检测并测序。结果 6例rpoB基因套式PCR扩增阳性者（其中4你为Bactec460培养阴性者），经DNA测序除1例rpoB基因无突变外，另5例均存在突变，3例为氨基酸密码子的第499位和第522位至第524位同时出现有义突变，其中第499位的基因突变为首次发现。结论 rpoB基因套式PCR检测在可辅助诊断结核病的同时，对其产物进行测序可直接判别对利福平耐药与否。     

Immunogenicity of recombinant BCG vaccine strains overexpressing components of the antigen 85 complex of Mycobacterium tuberculosis

The components of antigen 85 complex of Mycobacterium tuberculosis (Ag 85A, Ag 85B and Ag 85C), due to their immunodominant and secretory nature, represent promising protective antigen candidates and have been used in numerous vaccine preparations. We have used recombinant Bacille Calmette Guerin (BCG) strains overexpressing Ag 85A and Ag 85C to immunize BALB/c mice to investigate the immunogenicity of these strains. Mice immunized with recombinant BCG strains exhibited an increased humoral immune response when compared to mice immunized with wild-type BCG. The recombinant BCG strain overexpressing Ag 85A also induced an increased Th1-like response, characterized by elevated levels of IFN- in antigen stimulated splenocyte cultures and a strong IgG2a antibody response, when compared to wild-type BCG. Immunization with recombinant BCG strain overexpressing Ag 85C, on the other hand did not elicit increased IFN- secretion on restimulation of splenocytes in vitro.     

二甲基三十六烷基铵及卡介苗多糖核酸佐剂在结核亚单位疫苗加强BCG免疫中的效应研究

目的 探讨二甲基三十六烷基铵(dimo-thylidioctyl ammonium bromide,DDA)和卡介苗多糖核酸(BCC-PSN)佐剂在结核融合蛋白疫苗加强卡介苗(BCG)免疫中的不同免疫辅助效应.方法 选择DDA、BCG-PSN作为融合蛋白AMM(Ag85B-MPT64190-198-Mtb8.4)的佐剂,在BCG初免小鼠后,AMM疫苗加强免疫两次,其中一组联合使用两种佐剂(DDA/BCG-PSN),另一组单独以DDA作为佐剂,同时设立BCG或磷酸缓冲液(PBS)免疫组为对照.应用ELISA及ELISPOT检测免疫小鼠的体液与细胞免疫反应,最后一次加强免疫后第12周,以H37Rv尾静脉攻毒并检测小鼠肺脾组织细菌载量和病理改变,评价不同佐剂疫苗的保护效果.结果 在BCG初免基础上,联合佐剂组(AMM/DDA/BCG-PSN)和单独佐剂组(AMM/DDA)加强免疫两次后,脾脏淋巴细胞经抗原Ag85B和PPD(purified protein derivative)刺激后,皆可产生分泌较BCG组高的IFN-γ.毒力株攻击后菌落形成单位(colony-forming unit,CFU)计数显示,联合佐剂组(AMM/DDA/BCG-PSN)脾脏荷菌量少于PBS组和BCG组(P<0.05);而单独佐剂组(AMM/DDA)肺部荷菌量少于PBS组和BCG组(P<0.05).组织病理分析结果 表明AMM/DDA/BCG-PSN组肺组织病理损伤较轻,而AMM/DDA组病理损伤个体差异较大.结论 DDA是较为理想的结核亚单位疫苗佐剂,能诱导较强的细胞免疫和免疫保护作用;BCG-PSN可能具有免疫调节作用,可以减轻免疫病理损伤.     

结核分枝杆菌Ag85A质粒DNA疫苗治疗小鼠耐药结核病的效果研究

目的 研究结核分枝杆菌Ag85A质粒DNA疫苗治疗小鼠耐药结核病的效果.方法 用结核分枝杆菌高耐利福平低耐异烟肼临床分离株HB361尾静脉注射17～19 g的6～8周龄雌性BALB/c小鼠后,将小鼠随机均匀地分为6组,感染后第3天开始,分别用生理盐水(A组)、pVAX1空载体(B组)、利福平(C组)、微卡菌苗(D组)、Ag85A质粒DNA疫苗(E组)、利福平和Ag85A质粒DNA疫苗(F组)治疗60d,每组10只小鼠.治疗结束后3周,分别取肺和脾观察病理改变,称取重量做菌落计数.结果 治疗结束后3周,与对照组比较,D组、E组和F组肺脏病变有不同程度减轻,病变局限,病变范围分别为50%、20%、20%,2/3区域可见正常的肺泡结构,肺泡轮廓相对清晰,细胞分布均匀.与A组相比,D组、E组和F组肺脏菌落数分别减少了52%、68%、78%;脾脏菌落数依次减少了48%、65%、79%.结论 与对照组相比,Ag85A质粒DNA疫苗单独应用或与利福平联合应用治疗小鼠耐药结核病均显示疗效.     

Exposure to Mycobacterium avium primes the immune system of calves for vaccination with Mycobacterium bovis BCG

The objective of the investigation was to provide data on how a prior exposure of cattle to Mycobacterium avium, used here as a model of exposure to an environmental mycobacterium, affected the cellular immune response that follows vaccination with Mycobacterium bovis BCG. The assessment of cellular immune responses included lymphocyte proliferation assays, the delayed hypersensitivity skin test and IFN-gamma synthesis in whole blood cultures. One group of calves was inoculated subcutaneously with M. avium followed 12 weeks later by M. bovis-BCG. The other group was vaccinated subcutaneously with BCG alone. Calves previously exposed to M. avium responded more rapidly, as assessed in the in vitro assays, to purified protein derivative (PPD) from M. avium (PPD-A) or M. bovis (PPD-B) than did calves inoculated with BCG only, indicating that the exposure to M. avium had primed the immune response in these calves. Following inoculation of BCG the intensity of the in vitro responses and the delayed hypersensitivity skin test to PPD-A was higher for the M. avium-primed animals while the responses to PPD-B were similar in the M. avium-primed and BCG-only groups. The results are consistent with a model in which prior exposure to environmental mycobacteria does not necessarily inhibit the immune response to the vaccine strain, BCG. They suggest that M. avium infection primes the immune system of calves and that the detection of an immune response specific for M. bovis BCG is masked by reactivity to antigens also present in M. avium.     

Differentiation of clinical Mycobacterium tuberculosis complex isolates by their GyrB polymorphism

Purpose: To evaluate the reliability of the gyrB PCR-RFLP technique in differentiating clinical Mycobacterium tuberculosis complex isolates. Materials and Methods: A primer pair MTUB-f and MTUB-r for M. tuberculosis complex (MTBC) was used to differentiate 79 mycobacterial isolates by specific amplification of the 1,020-bp fragment of the gyrB gene (gyrB-PCR1). The MTBC isolates were further differentiated using a set of specific primers MTUB-756-Gf and MTUB-1450-Cr that allowed selective amplification of the gyrB fragment specific for M. tuberculosis (gyrB-PCR2). The DNA polymorphisms in the 1,020-bp gyrB fragment for 7 M. tuberculosis strains confirmed by PCR as well as 2 reference strains; M. tuberculosis H37Rv and M. bovis BCG were analyzed with the restriction enzyme Rsa1. Results: Seventy-seven (97.5%) isolates were positive for gyrB-PCR1 and thus identified as members of M. tuberculosis complex (MTBC) and two (2.6%) isolates were negative and identified as Mycobacteria other than tuberculosis (MOTT). All the M. tuberculosis isolates showed the typical M. tuberculosis specific Rsa1 RFLP patterns (100, 360, 560-bp) while 360 and 480-bp fragments were generated from M. bovis BCG. Conclusion: The gyrB PCR-RFLP using the endonuclease Rsa1 can be used to differentiate M. tuberculosis from M. bovis in clinical isolates.