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首页> 外文期刊>Infection and immunity >Characterization of Bartonella bacilliformis flagella and effect of antiflagellin antibodies on invasion of human erythrocytes.
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Characterization of Bartonella bacilliformis flagella and effect of antiflagellin antibodies on invasion of human erythrocytes.

机译：芽孢杆菌鞭毛的表征和抗鞭毛蛋白抗体对人红细胞的侵袭。

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Bartonella bacilliformis is the etiologic agent of Oroya fever in humans. Flagellum-mediated motility has been postulated as a major virulence factor for invasion of host cells. To address this hypothesis, we purified and characterized flagella from strain KC584 and then assessed their role in human erythrocyte association and invasion. Electron microscopy of the flagellar preparation showed a high concentration of filaments with a mean wavelength of 800 nm. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblot analysis, and KBr density gradient centrifugation indicated that the flagellar filament is composed of a polypeptide of 42 kDa. The flagellin is partially (ca. 50%) resistant to treatment with trypsin. The first 17 amino acid residues of the N terminus of the mature flagellin protein are GAAILTNDNAMDALQDL and show approximately 46% sequence identity to the residues of the N termini of two Caulobacter crescentus flagellin proteins. A monospecific polyclonal antibodies to the flagellin protein was generated, and its specificity was verified by both immunoblot and immunogold analyses. Human erythrocyte invasion assays performed with bartonellae exposed to the antiflagellin antiserum showed a significant decrease in bacterial association with and invasion of human erythrocytes in comparison with that in bartonellae exposed to preimmune rabbit serum or phosphate-buffered saline (PBS) controls. These results suggest that flagella are an important component in the invasiveness of B. bacilliformis.
机译：细菌杆菌巴尔通体是人类Oroya热的病原体。鞭毛介导的运动被认为是入侵宿主细胞的主要毒力因子。为了解决这个假设，我们从菌株KC584中纯化并鉴定了鞭毛，然后评估了它们在人红细胞缔合和侵袭中的作用。鞭毛制品的电子显微镜检查显示高浓度的细丝，平均波长为800 nm。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳，免疫印迹分析和KBr密度梯度离心表明，鞭毛丝由42 kDa的多肽组成。鞭毛蛋白对胰蛋白酶治疗具有部分抗性（约50％）。成熟鞭毛蛋白N末端的前17个氨基酸残基为GAAILTNDNAMDALQDL，与两个新月形鞭毛鞭毛蛋白N末端的残基具有约46％的序列同一性。产生了针对鞭毛蛋白的单特异性多克隆抗体，并通过免疫印迹和免疫金分析验证了其特异性。与暴露于免疫前的兔血清或磷酸盐缓冲盐水（PBS）对照的通心粉相比，暴露于抗鞭毛蛋白抗血清的通心粉进行的人红细胞侵袭测定显示与人红细胞的细菌缔合和侵袭显着降低。这些结果表明，鞭毛是芽孢杆菌侵袭性的重要组成部分。

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