• 伯氏疏螺旋体膜蛋白OspC研究进展 不要轻易放弃。学习成长的路上,我们长路漫漫,只因学无止境。


     莱姆病是由伯氏疏螺旋体惹起,即莱姆病螺旋体惹起的多零碎沾染性疾病。伯氏疏螺旋体自身并不具有生物活性,不克不及排泄毒力因子,其唯一的致毒因子为外膜脂卵白。此中,外膜脂卵白C(OspC)绝对外膜卵白A(OspA)以及鞭毛卵白来讲,其份子量较大。而且,最首要的一点是OspC在宿主体内能惹起晚期免疫反映;同时OspC 抗原性强,免疫OspC后能够

    呐喊

    呐喊检测出,血清中抗体的浓度较着回升。因而,本文对目前国内外对OspC的研讨举行了综述。?

    [要害词]伯氏疏螺旋体;莱姆病;OspC?

    [中图分类号] R514 [文献符号码] A [文章编号] 2095-0616(2015)02-26-04?

    Research progress of Borrelia burgdorferi on outer surface lipoprotein OspC protein?

    WANG Jiajie1 BAO Fukai1,2,3,4,5 LIU Aihua 2,3,4,5?

    1.Department of Microbiology and Immunology, The Institute for Tropical Medicine, Kunming Medical University, Kunming 650500, China; 2.Yunnan Province Key Laboratory for Tropical Infections Diseases in Universities, Kunming 650500, China;3. Yunnan Demonstration Base of International Science and Technology Cooperation for Tropical Diseases, Kunming 650500, China;4. Yunnan Province Integrative Innovation Center for Public Health, Diseases Prevention and Control, ?Kunming 650500, China; 5.Department of Biochemistry and Molecular Biology, Kunming Medical University , Kunming 650500, China?

    [Abstract] Lyme disease spirochaetes cause infectious of multiple system diseases. But Borrelia burgdorferi can not secrete toxins, its virulence factors mainly ?are outer surface lipoproteins (Osp). Among them, OspC can cause immune response early in the host. At the same time, OspC has strong immunogenicity, increased concentration of antibody in serum can be detected after immunized by OspC. The study progress of OspC was summarized here.?

    [Key words] Borrelia burgdorferi; Lyme disease; OspC?

    莱姆病(Lyme disease)作为一种蜱媒传布的传染性疾病, 在一些特殊人群和一些特别地域有很高的沾染率。但到目前为止,短少比拟迟钝和正确的诊断体式格局对莱姆病举行晚期诊断。因而,要做到实时防止和实时医治,在全国范围内仍是个困难。经研讨发觉,莱姆病的致病病原体是一种叫做伯氏疏螺旋体(Borrelia Burgdorferi)的螺旋体,其外膜卵白C(OspC)有十分强的抗原性和较强的免疫原性,机体在沾染莱姆病的晚期就能够

    呐喊

    呐喊发生特异性的IgM 抗体,将这类抗体纯化后,对植物模子举行免疫,了局发觉,植物模子在免疫后很短光阴内就可检测到高浓度的IgM 抗体,目前,重组OspC已用于莱姆病的临床诊断中。?

    1 OspC的细胞定位与化学布局?

    在举行OspC的抗原性研讨以前,起首对其细胞定位与化学布局做必然的研讨,才有可能片面的研讨其抗原性机制。?

    那末,外膜脂卵白C(OspC)是莱姆病螺旋体的次要外膜卵白之一,作为一种外膜卵白,OspC宽泛具有于螺旋体细胞外膜中。从布局上来看,β-桶状布局是其共有的布局特征,差别的β-桶状布局由差别的偶数个β-折叠片组成,β-折叠的数目从8 ~ 22个不等。在美国中西部、东北部等地域,发觉25种OspC基因型的表白[1-2]。Elisabeth Baum在其研讨中指出,OspC的179氨基酸残基至188个残基中的第五个C着末折叠布局是OspC的特异性交互抗体联合的要害地位[3]。OspC作为一个名义表露的脂卵白,能够

    呐喊

    呐喊惹起夙起的免疫反映[4-7]。在Elisabeth Baum的研讨中,得到了OspC的最大交互反映区域的3D图形。如图1,在图中,OspC二聚体的两条链别离被标以亮灰色与暗灰色。同时,在Elisabeth Baum的研讨中将OspC二聚体中与抗体药物关系度最高的区域指出。此中,绿色局部代表与种OspC有高关系度,黄色局部默示与两种OspC卵白有高关系度,红色局部默示与五种OspC卵白有高关系[8]。?

    图1 OspC的最大交互反映区域的3D图形?

    2 OspC的表白调控

    伯氏疏螺旋体特异性地调治其外膜脂卵白OspC的分解,以沾染宿主细胞。为了适应哺乳植物体内的免疫环境,螺旋体起首哄骗OspC在哺乳植物体内树立起其沾染过程。伯氏疏螺旋体大批发生OspA和OspB,而OspC在硬蜱体内低表白,从蜱前言传布到哺乳植物吸血后则涌现上调[9],以帮忙伯氏疏螺旋体转移到唾液腺[10],进而沾染哺乳类植物[11-12]。沾染一旦树立起来,OspC的表白就会被按捺,以防止被宿主的免疫零碎清除。同时伯氏疏螺旋体在人体内初期浮现一个高表白状态[13],当抗体发生后杀死了OspC一向涌现一个低表白克隆[14]。在表白基因的启动子中,反向反复序列的下游局部在调治OspC的表白中起到要害作用。Dan Drecktrah等[15]经由过程定点渐变技巧在份子程度上切割了OspC表白基因驾御子的着末反向反复序列,了局发觉,打乱着末反向反复序列但保存临近的反向反复序列能够

    呐喊

    呐喊

    呐喊阻止OspC的分解。同时发觉,温度,pH以及DNA超螺旋对OspC的分解也起到首要影响。Schwan等[16]发觉,在温度为32 ~ 37℃的条件下, OspC的表白量大幅度地进步。由此阐明

    顺叙,适合的温度是OspC 大批表白的首要条件, 在不适合的温度下,OspC不表白或表白量很低。因而,最近几年来良多国外学者以为,pH 值、温度以及其余要素或许会对OspC的免疫后果起到要害的影响[17-20]。更首要的是,比起核苷酸的序列,着末反向反复序列的碱基互补配对的具有对把持OspC的表白更为首要。Dan Drecktrah的了局显现,顺式作用元件在OspC毒力因子的表白上有要害的作用。?

    3 OspC致病性的研讨?

    在对伯氏疏螺旋体外名义膜卵白(OspC)致病性举行研讨时发觉,用OspC与Salp15联合,一同沾染哺乳类植物,能够

    呐喊

    呐喊帮忙伯氏疏螺旋体在局部假寓,并抵御免疫杀伤。2OspC作为一个无效的免疫靶,表白下调时帮忙伯氏疏螺旋体免疫逃逸,对OspC在晚期的沾染中起首要作用[13]。虽然OspC是伯氏疏螺旋体要害的毒力因子之一,但是它在伯氏疏螺旋体的沾染中的详细作用目前还不清楚。为了确定在接种了各种差别类型的OspC菌株后,OspC是否会惹起宿主反映, Antonara等别离接种了野生型OspC菌株,渐变型菌株以及齐全渐变型菌株后,对巨噬细胞,中性粒细胞以及细胞因子的发生举行了测定。在21种细胞因子的测定中发觉巨噬细胞趋化卵白(MCP-1),角质细胞来源趋化因子(KC, CXCL1)以及血管内皮生长因子(VEGF)在伯氏疏螺旋体的接种位点有表白量回升的趋势。而且在接种一周内的多个光阴点OspC的表白量有差别的变化[21]。?

    4 OspC的诊断代价?

    到目前为止,短少比拟迟钝和正确的诊断体式格局对莱姆病举行晚期的诊断。因而,要做到实时诊断和医治,在全国范围内仍是个困难。经研讨发觉,莱姆病的致病病原体伯氏疏螺旋体外膜卵白C(OspC)有较强的免疫原性,机体在沾染莱姆病的晚期就能够

    呐喊

    呐喊发生特异性的OspC IgM抗体,将OspC纯化后,对植物模子举行免疫,了局发觉,植物模子在免疫后很短光阴内就能够

    呐喊

    呐喊检测到高浓度的IgM抗体。目前,重组OspC已用于莱姆病的临床诊断。?

    最近几年的研讨表白,OspC作为伯氏疏螺旋体的次要外膜卵白之一,具有很强的抗原性。Gilmore等[17]哄骗OspC免疫12只初生小鼠, 然后哄骗伯氏疏螺旋体菌株沾染这12只取得免疫的初生小鼠, 由此发觉了OspC 能够

    呐喊

    呐喊

    呐喊使植物发生高浓度的抗体, 使其具有抵御伯氏疏螺旋体沾染的才能。?

    目前有不少研讨报道指出,OspC 在宿主体内能够

    呐喊

    呐喊惹起晚期免疫反映,因而有研讨者致力于哄骗OspC作为抗原用来诊断晚期的莱姆病[22-24]。早年,Hauser等[25]培养了大批差别莱姆病的菌株,并从中提取了大批的OspC作为抗原,哄骗ELASA技巧检测了222例莱姆病患者的血清和133例对照供血者的血清。其了局显现,有几株菌株间的IgG的检测活络度十分类似。哄骗Western blot技巧验证后发觉,OspC的确能够

    呐喊

    呐喊作为一种活络的莱姆病晚期诊断抗原。在此之后,Rousselle 等[6]经由过程对照772株广义氏疏螺旋体的OspC与14kD鞭毛卵白片断等其余细胞抗原平行使用于检测莱姆病病人和无症状的对照组的IgG与IgM, 了局发觉IgM抗体的差距性最大。别的, Hauser等[26]又做了进一步的研讨,哄骗WB技巧检测了菌株PKo、PBi与Pka2的活络度, 了局发觉OspC对晚期莱姆病沾染的检出活络度要远远高于其余抗原,并最终推荐了OspC作为PKo诊断的最迟钝的抗原。?

    Jobe等[27]的研讨中发觉,在人类莱姆病的晚期阶段中,ELISA能够

    呐喊

    呐喊作为一种无效的体式格局来检测OspC抗体,同时也能为胜利医治莱姆病作为一个首要的判断依据。?

    目前看来,OspC作为伯氏疏螺旋体的次要抗原之一, OspC虽然具有比拟大的差距, 但其具有很强的特异性。因而,在诊断学临床使用中,将OspC作为诊断抗原,具有很大的临床上风。同时,若是将OspC与其余的抗原相联合,举行配伍诊断,那末这类诊断体式格局不仅能够

    呐喊

    呐喊

    呐喊进步诊断的活络度, 还能淘汰与其余病原体的交叉反映。这将在未来莱姆病的诊断中显现出很强的上风与潜力。预计在今后的莱姆病防治中也会发挥必然的作用。至于在伯氏疏螺旋体的致病性上的研讨, 目前的研讨次要涉及于T细胞的免疫反映、病原体自身的作用以及差别基因型螺旋体的作用。预计在以后的研讨中,将会把OspC与其余卵白的研讨相联合,从而猎取更多的研讨了局,在莱姆病的各方面猎取进一步的希望。?

    付钰广等[28]哄骗来自SZ菌株的重组卵白OspC,举行WB实行检测显现,重组的OspC与阴性血清有较强的反映,同时SZ菌株作为我国的流行菌株,若哄骗SZ菌株的OspC树立绝对应的ELISA体式格局来检测伯氏疏螺旋体,这对我国莱姆病的防止和把持有很大作用,并有助于消除全国范围内莱姆病病患。同时,其研讨的重组卵白OspC(SZ菌株)为树立羊体内的伯氏疏螺旋体抗体的ELISA检测体式格局奠基了首要的基础。

    目前在全国范围内对OspC的研讨已有了初步了局,对OspC的研讨有助于对莱姆病的病发缘由,病发机理和病发规律举行理解。而且已初步树立了哄骗OspC作为检测抗原用来检测晚期的莱姆病。但对OspC的深入研讨还远远不敷,还有待更深一步的实行与临床研讨。?

    [参考文献]?

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