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首页> 外文学位 >Characterization of the pathogenesis of equine viral arteritis virus of cultured equine mononuclear phagocytes and pulmonary artery endothelial cells.
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Characterization of the pathogenesis of equine viral arteritis virus of cultured equine mononuclear phagocytes and pulmonary artery endothelial cells.

机译：培养的马单核吞噬细胞和肺动脉内皮细胞的马病毒性动脉炎病毒的发病机理的表征。

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摘要

Equine arteritis virus (EAV) infects endothelial cells (ECs) and macrophages in horses, and many of the clinical manifestations of equine viral arteritis (EVA) reflect vascular injury. Although there is only one serotype of EAV, there is marked variation in the virulence of different strains of the virus. In an attempt to determine if the in vitro growth characteristics of EAV are predictive of their virulence to horses, purified equine pulmonary artery endothelial cells (ECs) and mononuclear phagocytes were established and used as a model for the in vitro characterization of infection of equine cells with different strains of EAV. Initially, the replication and cytopathogenicity of three well-characterized strains of EAV of different virulence to horses were compared in rabbit kidney (RK-13) and ECs. Viral protein expression, plaque size, and cytopathogenicity of all three viruses were similar in RK-13 cells, whereas two virulent strains of EAV were readily distinguished from an avirulent strain by their plaque morphology and cytopathogenicity in primary equine ECs. EAV nucleocapsid protein was detected by flow cytometric analysis significantly later in ECs infected with the avirulent as compared to the virulent strains of EAV. Eleven additional strains of EAV of different virulence to horses were then tested in ECs and RK-13 cells to determine if the growth characteristics of this more extensive panel of EAV strains were also predictive of their virulence to horses. Whereas the growth characteristics of all these 11 strains of EAV were similar in RK-13 cells, virulent, and avirulent strains collectively were distinguished by their growth in ECs. Thus, primary equine ECs provide a convenient and relevant model for in vitro characterization of the pathogenesis of EVA and the virulence determinants of EAV.; To further characterize the mechanism of EAV-induced vascular injury and the potential role of virus-induced, macrophage-derived cytokines therein, cultured equine alveolar macrophages (AMΦ), blood monocyte-derived macrophages (BMΦ), and ECs were infected with either virulent (KY84) or avirulent (CA95) strains of EAV. Our data shows that the virulent KY84 strain of EAV induced significantly higher levels of mRNA encoding proinflammatory cytokines in infected AMΦ and BMΦ than did the avirulent CA95 strain. Treatment of equine ECs with the culture supernatant of EAV-infected AMΦ and BMΦ resulted in EC activation with cell surface expression of E-selectin, whereas infection of ECs with purified EAV caused only minimal expression of E-selectin. Specifically, the supernatant of KY84 EAV-infected AMΦ and BMΦ induced substantially higher levels of expression of E-selectin on ECs than did the supernatant of CA95 EAV-infected AMΦ and BMΦ. Moreover, the virulent KY84 strain of EAV induced significantly more TNF-α in AMΦ, BMΦ, and ECs than did the avirulent CA95 strain. Thus, EAV-induced EC- and macrophage-derived cytokines may contribute to the expression of EVA in horses, and the magnitude of the cytokine response and growth characteristics in equine mononuclear phagocytes and ECs reflects the virulence of the infecting virus strain. Collectively, equine ECs and mononuclear phagocytes provide a relevant and useful model to further characterize determinants of virulence and/or attenuation amongst strains of EAV.
机译：马动脉炎病毒（EAV）感染马的内皮细胞（EC）和巨噬细胞，马病毒性动脉炎（EVA）的许多临床表现都反映出血管损伤。尽管只有一种血清型的EAV，但不同病毒株的毒力存在明显差异。为了确定EAV的体外生长特征是否可预测其对马的毒力，建立了纯化的马肺动脉内皮细胞（EC）和单核吞噬细胞，并将其用作模型的模型。不同EAV株对马细胞感染的体外表征最初，在兔肾（RK-13​​）和EC中比较了三种对马有不同毒力的EAV毒株的复制和致病性。在RK-13​​细胞中，这三种病毒的病毒蛋白表达，噬菌斑大小和细胞致病性相似，而两种EAV毒株通过其斑马形态和在初级马EC中的细胞致病性很容易与无毒菌株区分开。通过流式细胞术分析，与无毒力的EAV菌株相比，在感染了无毒力的EC中，通过流式细胞术分析显着检测了EAV核衣壳蛋白。然后在EC和RK-13​​细胞中测试另外11种对马有不同毒力的EAV毒株，以确定这种更广泛的EAV毒株的生长特征是否也预示了它们对马的毒力。尽管所有这11种EAV菌株在RK-13​​细胞中的生长特征都相似，但有毒力和无毒力的菌株通过在ECs中的生长来共同区分。因此，初级马EC为EVA的发病机理和EAV的毒性决定因素的体外表征提供了方便且相关的模型。为了进一步表征EAV诱导的血管损伤的机制以及其中病毒诱导的巨噬细胞衍生的细胞因子的潜在作用，将培养的马肺泡巨噬细胞（AMΦ），血单核细胞衍生的巨噬细胞（BMΦ）和EC感染了两种（KY84）或无毒（CA95）的EAV株。我们的数据表明，与无毒力的CA95株相比，EAV的强毒KY84株在感染的AMΦ和BMΦ中诱导编码促炎细胞因子的mRNA水平显着提高。用被EAV感染的AMΦ和BMΦ的培养上清液处理马EC，导致E-选择素细胞表面表达引起EC活化，而纯化的EAV感染EC则仅引起E-选择素最低表达。具体而言，与CA95 EAV感染的AMΦ和BMΦ的上清液相比，KY84 EAV感染的AMΦ和BMΦ的上清液在EC上诱导的E-选择素表达水平高得多。此外，与无毒力的CA95株相比，EAV的强毒KY84株在AMΦ，BMΦ和EC中诱导的TNF-α明显更多。因此，EAV诱导的EC和巨噬细胞源性细胞因子可能有助于EVA在马中的表达，马单核吞噬细胞和EC中细胞因子反应的大小和生长特征反映了感染病毒株的毒力。集体地，马EC和单核吞噬细胞提供了相关且有用的模型，以进一步表征EAV毒株中毒力和/或减毒的决定因素。

著录项

  • 作者

    Moore, Brian David.;

  • 作者单位

    University of California, Davis.;

  • 授予单位 University of California, Davis.;
  • 学科 Biology Microbiology.; Biology Veterinary Science.; Agriculture Animal Pathology.
  • 学位 Ph.D.
  • 年度 2003
  • 页码 p.4758
  • 总页数 232
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 微生物学;
  • 关键词

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