title: Intracellular survival of Haemophilus somnus in bovine blood monocytes and alveolar macrophages.
abstract: The mechanisms used by Haemophilus somnus to survive and multiply within bovine mononuclear phagocytes are not fully understood. In order to study the interaction between bovine mononuclear phagocytes and H. somnus, a colorimetric assay using 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenylItetrazolium bromide (MTT) was developed to assess the survival of H. somnus within cultured bovine blood monocytes (BBM). Using this system, it was found that H. somnus was able to survive within BMM in vitro, and the kinetics of its survival were similar to that seen in BBM isolated from experimentally infected cattle. Using ultrastructural studies, it was possible to demonstrate the survival of H. somnus in freshly isolated bovine mononuclear phagocytes in membrane-bound vacuoles. To determine if activation of macrophage function would result in elimination of intracellular H. somnus, BBM were treated with E. coli lipopolysaccharide (LPS) or recombinant bovine (rBo) cytokines, interferon-gamma (IFN-gamma), granulocyte macrophage colony stimulating factor (GM-CSF), tumour necrosis factor-alpha (TNF-alpha) or interleukin-1beta (IL-1beta). Treatment of BBM with rBoIFN-gamma, rBoGM-CSF or E. coli LPS resulted in decreased intracellular survival of H. somnus at 18 and 48 h, whereas BBM treated with rBoTNF-alpha or rBoIL-1beta had reduced intracellular survival of H. somnus only at 18 h. However, none of these treatments resulted in complete elimination of the intracellular bacteria. The ability of H. somnus to survive and multiply in both freshly isolated and cytokine-treated cultured BBM demonstrated the capability of H. somnus to escape from macrophage killing mechanisms. This capability may play a role in the dissemination of H. somnus infection in the body.
authors.affiliation: Veterinary Infectious Disease Organization, 120 Veterinary Road, Saskatoon, Saskatchewan, S7N 5E3, Canada.
authors.forename: S M
authors.initials: SM
authors.lastname: Gomis
authors.forename: D L
authors.initials: DL
authors.lastname: Godson
authors.forename: G A
authors.initials: GA
authors.lastname: Wobeser
authors.forename: A A
authors.initials: AA
authors.lastname: Potter
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000276:immunology / Q000382:microbiology*; D003124:Colorimetry; D004396:Coloring Agents / Q000737:chemistry; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006190:Haemophilus / Q000254:growth & development* / Q000276:immunology / Q000502:physiology; D006192:Haemophilus Infections / Q000276:immunology / Q000382:microbiology / Q000662:veterinary*; D007371:Interferon-gamma / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D007700:Kinetics; D008070:Lipopolysaccharides / Q000494:pharmacology; D016676:Macrophages, Alveolar / Q000276:immunology / Q000382:microbiology* / Q000648:ultrastructure; D009000:Monocytes / Q000276:immunology / Q000382:microbiology* / Q000648:ultrastructure; D011994:Recombinant Proteins / Q000494:pharmacology; D013778:Tetrazolium Salts / Q000737:chemistry; D013844:Thiazoles / Q000737:chemistry; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D004396:Coloring Agents; D007375:Interleukin-1; D008070:Lipopolysaccharides; D011994:Recombinant Proteins; D013778:Tetrazolium Salts; D013844:Thiazoles; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; C022616:thiazolyl blue