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Francisella Tularensis infection of macorphage-like cell line J774.2 - changes of surface and intracellular molecules
Novosad, Jakub ; Krčmová, Irena (advisor) ; Raška, Milan (referee) ; Kročová, Zuzana (referee)
Summary: Background: Francisella tularensis (F.t.) is a facultative intracellular bacteria, enrolled at the list of Centre for Disease Control (CDC) as a high risk bioterrorism agent, category A. There is a long-term effort to understand to the immunopathogenesis of F.t. infection. The aim of our study was focused on phenotype analysis (CD54, CD16/32 and CD86) and nitric oxide (NO) production analysis of murine macrophage-like cell line J774.2 durig F.t. live vaccine strain (LVS) in vitro infection. J774.2 cells were either untreated or stimulated either before or after F.t. infection by interferon gamma (IFNγ), lipopolysaccharide (LPS) separately or in its combination. Method: We followed up the expression of cell surface markers and NO production 3, 6, 9, 12 and 24 hours after initiation of infection with or without stimulation using flow cytometry and Griess method respectively. The expression was followed as either absolute value of mean fluorescence index (MFI) or as relative change of MFI (∆MFI). Murine macrophage-like cells (J774.2) were incubated in cultivation flasks (2x106 cells/10ml of medium Dulbecco's MEM with Glutamax-1 with 10% BSA). The cells were activated with 10 or 50 ng of LPS / 1ml of medium or with 100 or 1 000 I.U. of IFNγ / 1ml of medium separately or in combination (10 ng/ml and 100...
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