Intracellular growth and survival of Vibrio cholerae in human macrophages
Abstract number: P1006
Objectives:Vibrio cholerae is Gram-negative bacteria found in water and it can be carried by sea living animals, such as shellfishes. V. cholerae O1 and V. cholerae O139 produce cholera toxin and cause cholera in humans. Despite that cholera infects millions and kills many thousands patients under pandemics and thus causes a global health problem, there is at present not enough information about how this bacterium lives in human. The prevalence rate of infections caused by Vibrio appears to be increasing globally. The combination of increased water temperature and salinity may contribute to increased association rates of the bacteria with sea living animals or protozoa. Author doctoral thesis showed that V. cholerae O1 and O139 grew and survived inside the protozoa Acanthamoeba castellanii. The current project is financed by the Swedish Civil Contingencies Agency (MSB), and is aimed to study abilities of clinical isolates of V. cholerae O1 and O139 to grow/survive inside human macrophages and disclose roles of bacterial capsule, lipopolysaccaride O-side chain and outer membrane protein A on the intracellular growth and survival of V. cholerae.
Methods and Microorganisms:Vibrio cholerae O1clasical, V. cholerae El Tor wild type, V. cholerae El Tor outer membrane mutant, V. cholerae O139 wild type, V. cholerae O139 capsule mutant, V. cholerae O139 capsule/LPS O-side chain double mutant, human macrophage TPH-1and human macrophage U937. An antibiotic assay used to differentiate between extracellular and intracellular V. cholerae. Vibrio species cultivated with human macrophages. Gentamicin used to kill extracellular bacteria. Intracellular growth and survival examined by viable count, fluorescent microscopy and RNA detection. Intracellular localisering visualized by confocal- and electron microscopy. Roles of bacterial capsule, lipopolysaccaride O-side chain and outer membrane protein A on the intracellular growth and survival of V. cholerae examined by statistical verified comparison between growth of wild type and mutant corresponded strain.
Results: The utilized methods showed that V. cholerae grew and survived inside human macrophages. The antibiotic assay differentiated between the extracellular and intracellular V. cholerae.
Conclusions: The intracellular behavior of V. cholerae may induce cell mediated and humoral immunity and explains complexity of V. cholerae and aids to new strategies for vaccination as well as treatment against cholera.
|Session name:||Abstracts of 21st ECCMID / 27th ICC|
|Location:||Milan, Italy, 7 - 10 May 2011|
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