| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2004: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2003: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Research Abstract |
Anaplama phagocytophilum and Ehrlichia chaffeensis are obligatory intracellular bacteria, agents of emerging infectious diseases (human granulocytic anaplasmosis and human monocytic ehrlichiosis), which infect professional phagocytes with tropism for granulocytes and monocytes/macrophages, respectively, and both are maintained with human promyelocytic cell lines HL-60 in many laboratories. Proteomic approach is a powerful technique to analyze a global protein expression profile in several cellular events of mammalian cell lines and tissues, plants, or bacteria. In this study, we applied to characterize the protein expression profile in host cell response to infection with A. phgocytophiluma and E. chaffeensis by using the most recently introduced DIGE (fluorescence two-dimensional difference gel electrophoresis) system (GE Healthcare, Piscatawy, NJ) which allowed us to detect differences in protein expression between infected and uninfected cells in a single gel. Proteomic changes were monitored in a time course on day 1, 2, and 3 postinfection and this approach revealed a total of 286 protein spots with abundance changes between infected and uninfected cells on day 3 postinfection. Using MALDI TOF/TOF tandem mass spectrometry, we successfully identified the protein species of 154 spots derived from HL-60 cells in response to infections with A. phagocytophilum and E. chafeensis. These protein species are associated with cytoskeleton, signal transduction, translation, stress response and more. MetaCore Pathway analysis (GeneGo Inc., St. Joseph, MI) suggested the candidate proteins of host cells associated with A. phagocytophilum and E. chaffeensis infections, probably these bacteria may reorganize host cell cytoskeleton through the protein-protein interaction to acquire their niches for intracellular survival.
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