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Advancements in Mammalian Comparative Immunology- Developing New Tools and Investigating the Innate Immune Response to Cedar Virus

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[Technical Report, Doctoral Thesis]

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Emerging viruses pose a serious threat to public health. Virtually all of the recent emerging virus outbreaks have come from human-wildlife interactions. These wildlife animals that harbor a virus are referred to as reservoirs. When the same virus then enters a human host however, overwhelming pathogenesis is often observed. Bats are the reservoirs for many of these zoonotic viruses. For example, Pteropus sp bats are the source of Hendra HeV and Nipah virus NiV. Both of these viruses cause severe disease with high mortality rates that are often a result of immunopathogenesis. Cedar virus CedV belongs to the same genus as HeV and NiV but is non-pathogenic. Here, CedV is used as a model for HeV and NiV to study human-bat comparative immunology. Because bats are the natural reservoirs of so many viruses, it is important to investigate how they are able to tolerate virus infections. A major goal of the present research is to develop a new technology that could allow a comparison of the human immune response to that of the viral reservoir. We use lentiviral transduction of two viral oncogenes, v-Raf and v-Myc, to immortalize macrophages isolated from murine blood. The resulting cell lines exhibit surface markers consistent with mature murine macrophages, most notably CD11b and F480. These cells are IFN responsive and capable of phagocytosis. Further, the cell lines are cytokine independent, which is important when working with species for which commercially available cytokines do not exist. We also investigate the NF-B response to CedV. This pathway is activated during many types of infections and is responsible for the subsequent activation of manypro-inflammatory genes. To compare the NF-B response in P. alecto reservoir to that of humans host we use kidney fibroblast cell lines from P. alecto PaKi and PaKiT and the human cell lines HEK 293T and MRC-5.

Subject Categories:

  • Microbiology

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[A, Approved For Public Release]