>> *To: The George Mason University Community*
>>
>> *Anne B. Verhoeven
>> PhD Biosciences Candidate
>> *
>> *Date: Thursday June 17, 2010
>> Time: 10:30 a.m.
>> Place: George Mason University
>> ** Room 256, Bull Run Hall
>> Prince William campus <http://www.gmu.edu/resources/visitors/findex.html>
>>
>> Dissertation Chair: Dr. Monique van Hoek
>> Committee members: Dr. Emanuel Petricoin III, Dr. Yuntao Wu, Dr. Timothy Born*
>>
>> *Title: "An Analysis of Early Signaling Pathways Activated by /Francisella Tularensis/ Infection"*
>>
>> A copy of the dissertation is on reserve in the Johnson Center
>> Library, Fairfax campus. The doctoral project will not be read at
>> the meeting, but should be read in advance.
>>
>> All members of the George Mason University community are invited to
>> attend.*
>> *
>>
>> *ABSTRACT:
>> *
>> /Francisella tularensis /is a zoonotic, gram negative pathogen that
>> in recent years has become a pathogen of increasing interest because
>> of its bioterrorism implications. /Francisella /has been classified
>> as a Type A pathogen by the CDC. It is capable of producing severe
>> infection with doses as low as 10 organisms. The possible threat of
>> /Francisella /as a biological weapon makes understanding its specific
>> pathogenicity of utmost importance.
>> /Francisella /is an intracellular pathogen that enters the cell
>> through an unknown method. Infection from /Francisella /occurs in
>> many different cell types, including macrophages, hepatocytes, and
>> endothelial cells. Once in the cell, /Francisella /is encapsulated
>> in a phagosome. Between 2-4 hours after the initial infection
>> /Francisella /escapes the phagosome and proceeds to replicate in the
>> hosts cytosol. Previous studies have demonstrated that /Francisella
>> /inhibits the release of pro-inflammatory cytokines such as
>> TNF-alpha, IL-1, IL-12, and IL-8, thus hampering the ability of the
>> innate immune system to respond to infection. Also, there is an
>> increase of IL-10 seen during /Francisella /infection. IL-10 is an
>> anti-inflammatory cytokine that further decreases the innate immune
>> system response. The lack of proper cytokine production might be
>> caused by /Francisella/'s ability to block the TLR pathways, CR3
>> pathways, and the apoptosis pathways, further impeding the innate
>> immune response. Interestingly, /Francisella /does not employ any
>> known secretion systems, such as type III or type IV secretion
>> systems that are commonly found in pathogenic bacteria. Nor does it
>> produce toxins that could explain its ability to block cell signaling
>> pathways. The mechanism used by /Francisella /to directly interact
>> with host cell pathways is still unknown.
>> *We hypothesize that by looking at specific pathways that are
>> activated during infection of J774A.1 macrophages by /Francisella
>> tularensis /LVS and /Francisella novicida, /we will be able to obtain
>> a phosphorylation map of activated host cell responses which could
>> lead to novel therapeutics.* The steps that we will use to accomplish
>> this task are: 1) the use of Reverse Phase Protein Microarray
>> analysis to generate a phosphorylation map of /Francisella /LVS and
>> /Francisella novicida/ infection in J774A.1 cells, 2) validation of
>> this map through western blots and analysis of previous reported data
>> on specific signaling pathways, 3) comparison of different
>> phosphorylation maps of /Francisella novicida /and /Francisella /LVS
>> to determine differences between the strains, and 4) in a separate
>> study, we will examing changes in gene expression in human lung
>> epithelial cells through the use of microarrays to determine the
>> difference in host cell gene transcription between uninfected A549
>> cells and /Francisella /LVS infected A549 cells.
>>
>>
>> ###
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