Bashmakov Yuriy K, Zigangirova Nailia A, Gintzburg Alexander L, Bortsov Petr A, Petyaev Ivan M
Yuriy K Bashmakov, Ivan M Petyaev, Cambridge Theranostics Ltd, Babraham Research Campus, Babraham, Cambridge, CB2 4AT, United Kingdom.
World J Hepatol. 2010 Feb 27;2(2):74-80. doi: 10.4254/wjh.v2.i2.74.
To evaluate the direct binding of two main chlamydial biovars (C. trachomatis and C. pneumoniae) to plasma lipoproteins and its effect on chlamydial infection rate in human hepatoma cell line (HepG2 cells).
Murine plasma lipoproteins were fractionated and isolated using fast-performance liquid chromatography (FPLC), spotted on nitrocellulose membrane and incubated with chlamydial suspensions. Direct binding of chlamydial particles to lipoprotein fractions has been studied using lipopolysaccharide-specific antibodies in immuno-dot blot binding assay and immunoprecipitation analysis. Immunostaining protocol as well as flow cytometry analysis have been employed to study the infectivity rate of chlamydial species in HepG2 cells.
Elementary bodies of both C. trachomatis and C. pneumoniae bind ApoB-containing fractions of plasma lipoproteins. That binding becomes stronger when heat-denatured FPLC fractions are used, suggesting a primary role of apolipoproteins in interaction between chlamydial particle and lipoprotein. Both chlamydial biovars efficiently propagate in human hepatoma cell line - HepG2 cells even in serum free conditions forming late-stage inclusion bodies and releasing extracellular elementary bodies. Preincubation of C. trachomatis and C. pneumoniae with native ApoB-containing lipoproteins enhances the rate of chlamydial infection in HepG2 cells.
A productive infection caused by C. trachomatis and C. pneumoniae may take place in human-derived hepatocytes revealing hepatic cells as possible target in chlamydial infection. Obtained results may suggest the participation of lipoprotein receptors in the mechanism of attachment and/or entry of chlamydial particles into target cells.
评估两种主要衣原体生物变种（沙眼衣原体和肺炎衣原体）与血浆脂蛋白的直接结合及其对人肝癌细胞系（HepG2细胞）中衣原体感染率的影响。
使用快速高效液相色谱法（FPLC）对小鼠血浆脂蛋白进行分级分离，点样于硝酸纤维素膜上并与衣原体悬液孵育。在免疫斑点印迹结合试验和免疫沉淀分析中，使用脂多糖特异性抗体研究衣原体颗粒与脂蛋白组分的直接结合。采用免疫染色方案以及流式细胞术分析来研究衣原体在HepG2细胞中的感染率。
沙眼衣原体和肺炎衣原体的原体均与血浆脂蛋白中含载脂蛋白B的组分结合。当使用热变性的FPLC组分时，这种结合会更强，表明载脂蛋白在衣原体颗粒与脂蛋白相互作用中起主要作用。即使在无血清条件下，两种衣原体生物变种都能在人肝癌细胞系 - HepG2细胞中有效繁殖，形成晚期包涵体并释放细胞外原体。沙眼衣原体和肺炎衣原体与天然含载脂蛋白B的脂蛋白预孵育可提高其在HepG2细胞中的感染率。
沙眼衣原体和肺炎衣原体引起的有效感染可能发生在人源肝细胞中，表明肝细胞可能是衣原体感染的靶标。所得结果可能提示脂蛋白受体参与衣原体颗粒附着和/或进入靶细胞的机制。
