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Ag85B

Ag85B的相关文献在2001年到2022年内共计68篇,主要集中在基础医学、分子生物学、肿瘤学 等领域,其中期刊论文68篇、专利文献3972篇;相关期刊45种,包括科学技术与工程、中国学术期刊文摘、现代生物医学进展等; Ag85B的相关文献由255位作者贡献,包括朱道银、江山、蒋英等。

Ag85B—发文量

期刊论文>

论文:68 占比:1.68%

专利文献>

论文:3972 占比:98.32%

总计:4040篇

Ag85B—发文趋势图

Ag85B

-研究学者

  • 朱道银
  • 江山
  • 蒋英
  • 陈全
  • 骆旭东
  • 师长宏
  • 杨晓峰
  • 杨登科
  • 赵勇
  • 靳风烁
  • 期刊论文
  • 专利文献

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    • 王丽梅; 姜泓; 康健; 靳芊芊
    • 摘要: 目的构建表达结核分枝杆菌多阶段抗原融合蛋白Ag85B-ESAT6(AE)和Rv2031c-Rv2626c(R2)的重组腺病毒,为其用于结核病新型疫苗的研究奠定基础。方法体外合成人延长因子1α(EF1α)启动子DNA序列,将其亚克隆至腺病毒穿梭载体pAdTrack-CMV中,构建含有CMV、EF1α双启动子的腺病毒穿梭载体。PCR扩增课题组前期构建的AE、R2融合蛋白基因,并依次亚克隆至上述腺病毒穿梭载体的CMV和EF1启动子下游,阳性重组穿梭质粒命名为pAd-AE-R2。将pAd-AE-R2与骨架质粒共转染HEK293细胞,包装获得重组腺病毒,命名为Ad-AE-R2。RT-PCR法和间接免疫荧光法(IFA)对重组腺病毒表达的AE、R2融合蛋白的基因和蛋白表达水平进行鉴定。结果Ad-AE-R2瞬时转染的HEK293细胞中可转录表达AE、R2融合蛋白基因。IFA法采用Ag85B、ESAT-6、Rv2626c和Rv2031c 4种抗原的单克隆抗体可分别检测到Ad-AE-R2感染的巨噬细胞中具有特异性的绿色荧光,表明Ad-AE-R2能够在宿主细胞内表达AE、R2融合蛋白。结论成功构建并包装获得表达结核分枝杆菌多阶段抗原融合蛋白AE、R2的重组腺病毒,为其用于结核病新型疫苗研究奠定基础。
    • 黄丹丹
    • 摘要: 目的:探讨Toll样受体4(TRL4)对Ag85B抗结核分枝杆菌DNA疫苗免疫保护效应的影响。方法:分别用pcDNA-Ag85B,pcDNA-Ag85BTRL4免疫小鼠,第三次免疫2周后检测细胞因子IFN-γ,IL-4和IL-10分泌水平,3月后进行MTB感染实验,观察MTB菌落数。结果:TRL4可增强Ag85B免疫后IFN-γ分泌量,IL-4和IL-10分泌水平无显著变化。免疫后小鼠抗MTB能力增强,pcDNA-Ag85B,pcDNA-Ag85B-TRL4组MTB菌落数均有减少且pcDNA-Ag85B-TRL4组抗MTB作用更明显。结论:TRL4可增强Ag85B抗MTB DNA疫苗免疫保护效应。
    • 方习静; 刘蓉娜; 张海峰; 段凯; 闭兰
    • 摘要: 目的:利用液质联用技术分析比较基因重组卡介苗Aeras-422与传统卡介苗的主要分泌型抗原Ag85复合体成分。方法分别提取基因重组卡介苗Aeras-422与传统卡介苗培养上清中的分泌蛋白,采用反相高效液相色谱(Reversed phase high performance liquid chromatography,RP-HPLC)进行分离,并将最终得到的目的蛋白峰采用基质辅助激光解吸电离飞行时间质谱仪( MALDI-TOF-MS ReflexⅢ)进行质谱分析鉴定。结果基因重组卡介苗Aeras-422与传统卡介苗培养上清分泌蛋白Ag85复合体成分有所不同,Aeras-422表达的Ag85复合体成分包括Ag85A和Ag85B两种抗原,传统卡介苗菌株仅分泌表达Ag85A抗原。结论基因重组卡介苗Aeras-422分泌Ag85复合体的能力优于传统卡介苗。为建立新型结核病疫苗的质控方法提供了数据支持。%Objective To establish a simple method for the determination and comparing the secretory antigen Ag85 protein complex components from two kinds of TB vaccine strains(AERAS-422 and Danish-SSI 1331 BCG vaccine).Methods Collecting the cultured supernatants of AERAS-422 and Danish-SSI 1331 BCG vaccine strains, the samples were concen-trated by ultrafiltration,then, analysed by reverse high performance liquid chromatography.Collecting the main peak to ana-lyse by SDS-PAGE, and the peaks of protein for mass spectrometric was determined using matrix assisted laser desorption/ionization time of flight mass spectrometry ( MALDI-TOF-MS ReflexⅢ) .Results By optimizing the reversed phase liquid chromatography separation process, the Ag85 mixture composition were further separated, and to establish Ag85A and Ag85B peak corresponding peptides mass spectrum maps by MALDI TOF-MS technology.The Ag85 complex has the differ-ent components in the cultured supernatants of two kinds of strain.Ag85 complex of Aeras-422 strain was composed with Ag85A and Ag85B.The traditional BCG vaccine strain only expressed the Ag85A antigen.Conclusion Establishing a simple and effective quality control method for new tuberculosis vaccine research.
    • 赵青; 冯国栋; 王雯菁; 朱旗; 张敏; 刘扬; 史明; 赵钢
    • 摘要: 目的 观察结核分枝杆菌及其抗原成分对人外周血中性粒细胞白三烯B4表达的影响. 方法 ELISA法检测结核分枝杆菌不同菌株及抗原分别与外周血中性粒细胞共培养上清白三烯B4及相关细胞因子TNF-α表达量,应用花生四烯酸代谢通路抑制剂来探讨其可能的机制. 结果 BCG、ESAT-6、Ag85B刺激中性粒细胞生成白三烯B4和TNF-α、ESAT-6、Ag85B刺激生成LTB4呈剂量依赖,H37Ra不引起LTB4表达但可增加TNF-α表达,Ag85B刺激生成的LTB4 可被齐留通和苯丁抑制素阻断,但对TNF-α表达无明显影响.结论 卡介苗及ESAT-6、Ag85B诱导人中性粒细胞产生LTB4和TNF-α,Ag85B诱导的LTB4可被抑制剂阻断但不影响TNF-α表达. 感染早期中性粒细胞通过生成LTB4发挥抗结核作用.%Objective To study the effects of Mtb and antigen on leutrinene B4 production from human neutrophil. Methods The fresh peripheral blood samples from healthy adults were incubated with Mtb and antigen at different dose. Supernatants were collected for LTB4 and TNF-α detection by ELISA. Pharmacological inhibitors of leukotrienes synthesis was used to find possible mechanism. Re-sults In this study we found that neutrophils could release LTB4 and TNF-αinduced by BCG,ESAT-6 and Ag85B. ESAT-6-and Ag85B-activated human PMN secreting the LTB4 in a dose dependent manner. Neutrophils did not produce LTB4 in response to H37Ra. Treat-ment of PMN with the leukotriene B4 inhibitor Zileuton or Bestatin prior to stimulation with Ag85B partially blocked LTB4 induction but did not work on TNF-α production. Conclusion BCG,ESAT-6,Ag85B may induce neutrophil activation and enhance LTB4 release. These effects can be reduced by arachidonic acid metabolism pathway inhibitors of Zileuton and Bestatin which did not work on TNF-αproduction. We conclude that PMN contribute to early resistance to Mtb via LTB4 secretion.
    • 杜昭弘; 胡东; 吴静; 张荣波; 王婉; 戴京京; 何江; 王文洋; 杨小康; 邢应如; 尤其; 穆敏
    • 摘要: Objective To construct pET28a-PTD-Ag85B prokaryotic expression vector, and to explore the optimization of expression conditions in E.coli BL21.Methods Using restriction enzymes Nhe I and BamH I to digest plasmid pET28a and then insert the TAT-PTD sequence into it to obtain pET28a-PTD recombinant plasmid, using RT-PCR for amplification of Ag85B gene, and then cloning to pET28a-PTD plasmid to obtain pET28a-PTD-Ag85B recombinant plasmid.Conversion pET28a-PTD-Ag85B recombinant plasmid to E.coli, isopropyl thiogalactoside ( IPTG ) induces PTD-Ag85B fusion protein expression, using polyacrylamide gel electrophoresis ( SDS-PAGE ) identify the expression product.Screening optimal inducer concentration, the optimum induction temperature and induction time, refolding inclusion by urea, separation and purification the fusion protein with a His-tag ion affinity chromatography.Results The recombinant plasmid pET28a-PTD-Ag85B was successfully constructed, could highly express PTD-Ag85B fusion protein in E.coli BL21 ( DE3 ). Conclusions Successful expression and purification the PTD-Ag85B proteins, in order to obtain a large number of PTD-Ag85B fusion protein provides technical reserves.%目的:构建pET28a-PTD-Ag85B原核表达载体,并对其在E.coli BL21(DE3)中表达条件进行优化。方法将TAT-PTD序列插入到用限制性内切酶Nhe I和BamH I双酶切的pET28a质粒中,获得pET28a-PTD重组质粒;RT-PCR法扩增Ag85B基因并将其克隆至pET28a-PTD质粒中,获得pET28a-PTD-Ag85B重组质粒。将重组质粒pET28a-PTD-Ag85B转化至大肠杆菌,异丙基硫代半乳糖苷( IPTG)诱导PTD-Ag85B融合蛋白表达,聚丙烯酰胺凝胶电泳( SDS-PAGE)鉴定表达产物。筛选最适诱导剂浓度、诱导温度和时间,尿素复性包涵体,并用His-tag离子亲和层析柱分离纯化融合蛋白。结果成功构建pET28a-PTD-Ag85B重组质粒,在E.coli BL21中能高效表达PTD-Ag85B融合蛋白。结论成功表达及纯化了PTD-Ag85B蛋白,为获得大量PTD-Ag85B融合蛋白提供了技术储备。
    • 李华; 李枫; 梁海燕; 黄艺; 米利古; 王仙; 李永祥; 曹帅丽; 申爱平
    • 摘要: 目的 比较结核分枝杆菌异烟肼耐药株与敏感株fbpB基因mRNA转录水平及蛋白水平表达的差异性.方法 利用比例法对分离出来的细菌做药敏分析及鉴定试验,然后提取细菌的总RNA及蛋白质.利用qRT-PCR技术检测异烟肼耐药株与敏感株fbpB基因mRNA水平的差异,并用两个独立样本的非参数检验进行统计分析,再利用Western blot技术检测其蛋白水平有无差并,两个独立样本的t检验作统计学分析.结果 结核分枝杆菌耐药株和敏感株mRNA水平表达量分别为0.80±0.20、0.66±0.12,蛋白相对于内参的表达量分别为0.96±0.12、0.53±0.08.耐药株在两个水平上均高于敏感株(P<0.05).结论 结核分枝杆菌对异烟肼产生耐药与fbpB基因编码的蛋白Ag85B有一定的相关性.
    • 梁艳; 吴雪琼; 阳幼荣; 张俊仙
    • 摘要: 目的 研究结核分枝杆菌Ag85A和Ag85B重组蛋白疫苗联合佐剂对结核的治疗作用.方法 用结核分枝杆菌H37Rv通过尾静脉注射60只BALB/c小鼠,分别用生理盐水、利福平、草分枝杆菌F.U.36注射液、佐剂77-1、Ag85A和Ag85B重组蛋白疫苗、Ag85A和Ag85B重组蛋白疫苗联合佐剂治疗.免疫结束后2周杀鼠,观察肺和脾病理改变,称取重量,做菌落计数.结果 肺组织病理显示:生理盐水组肺组织病变严重、广泛,佐剂组比生理盐水组病变略轻,病变仍较重、广泛,草分枝杆菌F.U.36注射液组、Ag85A和Ag85B重组蛋白疫苗组、Ag85A和Ag85B重组蛋白疫苗联合佐剂组肺组织病变减轻、局限,3/5区域肺泡结构完整、清晰,利福平组肺组织无病变.与生理盐水组比较,利福平组、草分枝杆菌F.U.36注射液组、Ag85A和Ag85B重组蛋白疫苗联合佐剂组肺脏菌落数分别减少2.11 log、0.76 log和0.57 log;肝脏菌落数分别减少2.11 log、0.74 log和0.44 log(t>3.21,P<0.01).结论 Ag85A和Ag85B重组蛋白疫苗联合佐剂对小鼠结核病具有较好的治疗效果,与草分枝杆菌F.U.36注射液相当.%Objective To evaluate the therapeutic effects of Ag85A and Ag85B recombination protein vaccines combined with adjuvant in mouse model of Mycobacterium tuberculosis infection to establish new immunotherapeutic agents to treat tuberculosis.Methods 60 female BALB/c mice were infected intravenously via the tail vein with Mycobacterium tuberculosis H37RV,then randomly divided into six groups based and treated as follow at the third day after infection:saline,rifampin,Mycobacterium Phlei F.U.36 injection,adjuvant,Ag85A and Ag85B recombinat protein vaccines,Ag85A and Ag85B recombinat protein vaccines combined with adjuvant.Vaccines were injected intramuscularly three times at two-week intervals.Mice were sacrificed two weeks after the final immunization.The lungs and spleens from the mice were taken and their pathological changes,weight and number of mycobacterial colony were examined.Results The histopathological changes of lung showed that the lung lesions were severe and extensive in saline and adjuvant group,the lung lesions were slight and limited,there were relatively clearand normal structure of alveoli in Mycobacterium Phlei F.U.36 injection,adjuvant,Ag85A and Ag85B recombinatprotein vaccines,Ag85A and Ag85B recombinat protein vaccines combined with adjuvant group,there were no lung lesions in rifampin group.Compared with saline group,pulmonary bacterial loads of rifampin group,Mycobacterium Phlei F.U.36 injection group,Ag85A and Ag85B recombinat protein vaccines combined with adjuvant group reduced by 2.11 log,0.76 log,and 0.57 log,liver bacterial loads by 2.11 log,0.74 log,and 0.44 log (t > 3.21,P < 0.01),respectively.Conclusions The immunotherapeutic effect of Ag85A and Ag85B recombinat protein vaccines combined with adjuvant is significant in the mouse model of tuberculosis,as that as Mycobacterium Phlei F.U.36 injection.
    • 杜久伟; 胡东; 吴静; 王文洋; 杨小康; 许礼发; 张荣波
    • 摘要: 目的 应用分子克隆技术获得纳米级Ag85B蛋白并对其进行鉴定和免疫学特性检测.方法 利用S-layer/Streptag I这一具有自我组装能力的纳米模式嵌合结核杆菌分泌的Ag85B蛋白基因,将其克隆到原核表达载体pET28a(+)中,并在大肠杆菌中表达Ag85B蛋白,表达后的蛋白经SDS-PAGE和Western blotting鉴定.结果 原核表达重组质粒pET28a-Streptag I-SbpA-Ag85B成功构建,并可在大肠杆菌中诱导表达,得到的Streptag I-Sbpa-Ag85B蛋白经SDS-PAGE和Western blotting鉴定正确,重组蛋白能被结核患者血清识别.结论 重组后的Ag85B蛋白具有较强的免疫反应性.所建立的原核表达体系为新型结核疫苗研制提供了一个新的思路.%Objective To obtain nanometer Ag85B protein by the molecular cloning techniques and identify its immunological characteristics.Methods The gene sllB coding surface layer protein (S-layer)was truncated and linked with the Mycobacterium tuberculosis secreted protein Ag85B gene.The expression plasmid pET28a-Streptag I-SbpA-Ag85B was transformed into E.coli,the protein was identified by SDS-PAGE and Western blotting.Results The expression system was developed successfully for recombinant nanometer-scale antigen and the fusion protein was identified by SDS-PAGE with correct molecular weight and can be recognized by the serum of tuberculosis patients.Conclusion The recombinant Ag85 B protein has strong immune reaction and small size on nanometer level,this expression system provides a new way for the tuberculosis vaccine design.
    • 陶成武; 赵丹; 董慧; 单发; 连凯; 潘志明; 陈祥; 殷月兰; 焦新安
    • 摘要: [目的]Rv1886c基因编码的Ag85B是结核分枝杆菌(Mycobacterium tuberculosis,M.tb)感染早期的分泌蛋白,本研究对其所诱导的免疫应答特性进行了探索.[方法]对Ag85B进行原核表达和鉴定,并通过夹心ELISA、间接ELISA及ELISPOT方法测定其诱导的细胞免疫和体液免疫应答水平.[结果]SDS-PAGE及Western blot鉴定结果表明,以包涵体形式表达的Ag85B蛋白,经变性、复性后能与结核病人的抗血清及免疫重组李斯特菌LM-Ag85B的小鼠抗血清发生特异性反应,表明His-Ag85B融合蛋白具有较好的免疫活性.将纯化的Ag85B蛋白皮下免疫C57 BL/6小鼠,夹心ELISA的测定结果表明,Ag85B蛋白免疫组诱导小鼠产生的特异性IFN-γ水平显著高于IL-4的水平(P <0.001),呈现Thl型细胞免疫应答趋势;以结核菌素PPD作为包被抗原,通过间接ELISA测定的血清抗体效价达到1:6400,表明Ag85B也能诱导有效的体液免疫应答.此外,以尾静脉途径初次免疫小鼠42天时,ELISPOT测定结果显示,结核分枝杆菌H37Rv诱导小鼠产生Ag85B240-259特异性的IFN-γ水平极显著高于卡介苗(BCG)免疫组(P<0.001).[结论]Ag85B蛋白能激发小鼠产生较强的Thl型细胞免疫应答和较好的体液免疫应答;BCG单次免疫后诱导小鼠产生的Ag85B特异的细胞免疫应答水平较低.本研究为揭示结核分枝杆菌的致病机理、新型疫苗的研制和早期诊断试剂的开发奠定了基础.
    • 曹德君; 陈颖钰; 郭爱珍; 陈焕春
    • 摘要: 结核病是一种慢性消耗性人畜共患病,不仅严重影响人类的身体健康,还对畜牧业造成惨重的经济损失.卡介苗是目前官方唯一认可使用的疫苗,但仅对儿童具有较好的保护力;部分国家也将卡介苗应用于动物结核病的预防中,但效果一般.本研究以小鼠为实验模型,对前期通过基因工程方法构建的增强型重组卡介苗GM-CSF-Ag85B-Rv3872-CFP10-ESAT6-BCG的免疫效果进行了评价,证实了该候选疫苗对小鼠的安全性,且能够产生针对BCG外源蛋白的特异性体液免疫和细胞免疫,对结核病具有一定的保护作用.
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