Burkholderia pseudomallei is a gram-negative opportunistic soil pathogen that causes the life-threatening disease melioidosis. It is endemic in Northern Australia and Southeast Asia but can be found throughout many other regions in the world. Diabetes mellitus is a predisposing risk factor for infection with this organism and it has been demonstrated that diabetic males are particularly susceptible to severe infection. Previous research suggested that monocytes isolated from the whole blood of diabetic males demonstrated a decreased ability to produce the proinflammatory cytokines IL-1β and IL-8. We hypothesized that monocyte-derived macrophages from diabetic males would also secrete lower levels of pro-inflammatory cytokines and that this difference between gender and diabetic state would be more pronounced compared to those seen previously with monocytes. Twenty volunteer with type I diabetes mellitus (ten males and ten females), along with twenty healthy age- and gender-matched controls donated blood for this study. Monocytes were collected from whole blood and allowed to differentiate into macrophages with the use of human recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF). Macrophages were then divided into groups and infected with B. pseudomallei, B. thailandensis (a closely related by non-pathogenic bacterium that inhabits similar niches), and E. coli. An uninfected control was used as well. At six hours post-infection, mRNA was collected from all cells and qPCR was performed to determine cytokine expression levels. All mRNA values collected from cells which had been infected with bacterial agents were normalized against the corresponding concentrations of mRNA from mock-infected cells. Mean log fold increases in both IL-1β and IL-8 were computed and compared. Preliminary testing showed decreased levels of both IL-1β and IL-8 from B. pseudomallei-infected macrophages isolated from a diabetic male compared to the healthy, age-matched male control. Surprisingly, results from all forty donors demonstrated that gender and diabetic state were not significant factors in the proinflammatory responses of macrophages infected with B. pseudomallei, although further testing is needed to determine if these results were influenced by experimental parameters.
| Identifer | oai:union.ndltd.org:BGMYU2/oai:scholarsarchive.byu.edu:etd-3645 |
| Date | 17 November 2010 |
| Creators | Blam, Annette J. |
| Publisher | BYU ScholarsArchive |
| Source Sets | Brigham Young University |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Theses and Dissertations |
| Rights | http://lib.byu.edu/about/copyright/ |
Page generated in 0.0023 seconds