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Mammalian host cell reservoirs during Anaplasma infection.

机译：无浆膜感染期间的哺乳动物宿主细胞库。

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摘要

Endothelial cell culture and preliminary immunofluorescent staining of Anaplasma-infected tissues suggest that endothelial cells may be an in vivo nidus of mammalian infection. To investigate endothelial cells and other potential sites of Anaplasma infection in mammalian tissues, a sensitive and specific, in situ rolling-circle amplification technique to detect localized Anaplasma gene sequences was developed. Via the technique described here and von Willebrand factor immunofluorescence, A. phagocytophilum and A. marginale were successfully localized in situ within cultured mammalian cells. This is the first application of this in situ method for detection of a microorganism and forms the foundation for applications of this technique to detect, localize, and analyze Anaplasma nucleotide sequences in the tissues of infected hosts and in cell cultures.;Three Anaplasma-infection trials using immunocompetent dogs and cattle were performed to investigate different aspects of endothelial cells as they relate to Anaplasma life cycles and to further describe clinical aspects of Anaplasma pathogenesis. Four Beagles were inoculated with A. phagocytophilum from different sources, allowed to develop chronic infection, and treated with doxycycline. Regardless of isolate or duration of doxycycline treatment, A. phagocytophilum DNA remained detectable for several months in blood and tissues, though 8 organisms were not identified microscopically. This is the first infection of dogs using cultured endothelial cells as the source of inoculum, the first demonstration of molecular evidence of chronic, persistent infection in blood and tissues of subclinical dogs despite doxycycline treatment, and the first investigation of endothelial cells as a potential in vivo source of A. phagocytophilum during chronic canine infection using in situ rolling-circle amplification. Two steers were inoculated with A. marginale by tick-feeding transmission and were euthanized at different points within the parasitemic cycle. The tissue distribution of A. marginale during peak and trough parasitemia was described using real-time PCR, though organisms were not identified in tissues microscopically. This is the first survey of A. marginale tissue distribution after tick-transmission and the first investigation in immunocompetent cattle of endothelial cells as a potential in vivo source of A. marginale in tick-bite sites and distant tissues using three techniques (immunofluorescence, electron microscopy, in situ rolling-circle amplification).
机译：内皮细胞培养物和无瓣膜感染组织的初步免疫荧光染色表明，内皮细胞可能是哺乳动物感染的体内病原体。为了研究哺乳动物组织中内皮细胞和厌氧菌感染的其他潜在部位，开发了一种灵敏且特异的原位滚环扩增技术来检测局部厌氧菌基因序列。通过此处所述的技术和冯·威勒布兰德因子免疫荧光技术，吞噬链球菌和边缘拟杆菌被成功地原位定位在培养的哺乳动物细胞内。这是这种原位检测微生物的方法的首次应用，并为应用该技术检测，定位和分析被感染宿主组织和细胞培养物中的无核苷核苷酸序列奠定了基础。进行了具有免疫能力的狗和牛的试验，以研究内皮细胞与无生命浆体生命周期相关的不同方面，并进一步描述无生命浆体发病机理的临床方面。用来自不同来源的吞噬链球菌接种四只小猎犬，使其发生慢性感染，并用强力霉素治疗。不管分离多西环素或多西环素治疗的持续时间如何，尽管在显微镜下没有发现8种生物，但嗜血曲霉DNA在血液和组织中仍可检测数月。这是使用培养的内皮细胞作为接种物的犬的首次感染，尽管强力霉素治疗，但仍首次证明了亚临床犬血液和组织中慢性，持续感染的分子证据，并且首次研究了内皮细胞作为潜在的感染源。使用原位滚环扩增在慢性犬感染过程中吞噬链球菌的体内来源。通过tick饲传播将两头ers牛接种了拟南芥，并在寄生虫周期的不同时间点对其实施了安乐死。尽管没有在显微镜下鉴定出生物体，但使用实时PCR描述了在峰谷和谷底寄生虫病期间边缘农杆菌的组织分布。这是tick传播后首次对角缘拟南芥组织分布的调查，也是对使用三种技术（免疫荧光，电子技术）在tick咬位点和远处组织中具有免疫力的牛作为内皮tick的潜在体内来源的首次调查。显微镜，原位滚环扩增）。

著录项

  • 作者

    Wamsley, Heather L.;

  • 作者单位

    University of Florida.;

  • 授予单位 University of Florida.;
  • 学科 Biology Molecular.;Biology Veterinary Science.;Biology Microbiology.
  • 学位 Ph.D.
  • 年度 2009
  • 页码 92 p.
  • 总页数 92
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分子遗传学;微生物学;动物学;
  • 关键词

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