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首页> 外文期刊>Infection and immunity >Alp, an Arthropod-Associated Outer Membrane Protein of Borrelia Species That Cause Relapsing Fever
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Alp, an Arthropod-Associated Outer Membrane Protein of Borrelia Species That Cause Relapsing Fever

机译：Alp，一种与节肢动物相关的疏螺旋体物种外膜蛋白，可引起复发性发热

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Borrelia hermsii and other relapsing fever (RF) species are noted for their highly polymorphic surface antigens, the variable major proteins (VMP). Less is known about other surface proteins of these pathogens in either their vertebrate reservoirs or arthropod vectors. To further characterize these proteins, we elicited antibodies against VMP-less cells, noted antibody reactions against whole cells and cell components, and then subjected selected antigens to mass spectroscopy for amino acid sequencing for comparison against a B. hermsii genome database. One of the derived monoclonal antibodies, H0120, agglutinated spirochetes, and in Western blot analyses, it bound to a 14-kDa protein of whole cells and their membrane fractions but not after protease treatment. A search of open reading frames of the B. hermsii genome with extracted peptides identified the 14-kDa protein with bha128, a 453-nucleotide gene of the 175-kb linear plasmid. The bha128 gene was synthesized and expressed in Escherichia coli. The protein product was bound by antibody H0120. Genes homologous to bha128 occur in the RF species Borrelia turicatae, B. duttonii, and B. recurrentis but not in Lyme disease Borrelia species or other organisms. The following findings indicated an association of BHA128, renamed Alp, with the tick environment: (i) Alp was produced at higher levels at 23°C than at 34°C; (ii) almost all spirochetes in tick salivary glands were bound by the H0120 antibody, but only ～1% of spirochetes in the blood of infected mice were bound; and (iii) infected mice produced antibodies to several B. hermsii antigens but not detectably to native or recombinant Alp.
机译：伯氏疏螺旋体和其他复发性发热（RF）物种因其高度多态的表面抗原（可变主要蛋白（VMP））而闻名。对于这些病原体的脊椎动物水库或节肢动物载体中其他表面蛋白的了解较少。为了进一步表征这些蛋白质，我们引出了针对无VMP的细胞的抗体，注意到了针对全细胞和细胞成分的抗体反应，然后对选定的抗原进行了质谱分析，以进行氨基酸测序，以便与B. hermsii基因组数据库进行比较。一种衍生的单克隆抗体，H0120，凝集的螺旋体，在Western印迹分析中，它与全细胞及其膜级分的14 kDa蛋白结合，但在蛋白酶处理后不结合。用提取的肽对布鲁氏杆菌基因组的开放阅读框进行搜索，鉴定出带有bha128的14-kDa蛋白，bha128是175-kb线性质粒的453个核苷酸的基因。合成了bha128基因并在大肠杆菌中表达。蛋白质产物与抗体H0120结合。与bha128同源的基因出现在RF物种伯氏疏螺旋体，duttonii和轮状芽孢杆菌中，而在莱姆病伯氏疏螺旋体物种或其他生物中不存在。以下发现表明，BHA128（重命名为Alp）与壁虱环境有关：（i）Alp在23°C的水平比在34°C的水平更高; （ii）tick唾液腺中几乎所有的螺旋体都被H0120抗体结合，但被感染小鼠血液中只有〜1％的螺旋体被结合。 （iii）感染的小鼠产生针对几种疱疹双歧杆菌抗原的抗体，但未检测到针对天然或重组Alp的抗体。

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