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This study investigated the role of autophagy in the survival of the invasive bacterium Brucella melitensis strain 16M in murine macrophages. Here, Brucella melitensis 16M was found to trigger autophagosome formation, enhance autophagy flux and increase the expression level of the autophagy marker protein LC3-II. When autophagy was pharmacologically inhibited by 3-methyladenine (3-MA), Brucella replication efficiency was significantly decreased (p < 0.05). These results suggest that autophagy favors Brucella melitensis 16M survival in murine macrophages.
Bacterial movement inside the cytoplasm is a major virulence factor in that it is necessary for efficient colonization of the infected tissues. Molecules from both the host and the pathogen present possible sites of pharmacologic intervention. Because locomoting Listeria and Shigella mimic the activated state of the leading edge of nonmuscle cells, these pathogens are powerful tools for dissecting the molecular machinery of actin-based motility. Analysis of the movement linked to cytoskeleton may lead to: (I) improved understanding of the mechanisms of disease transmission, including carriers and carrier states, pathogen movements, environmental factors and pharmacokinetics of the uptake and residues of vaccines and other biologies, and drugs in cultivated organisms; (II) new therapeutic developments, since it identifies the molecular targets involved in the pathogenicity of Listeria and Shigella and vaccinia intracellular enveloped virus. Recent knowledge about the intracellular movement in cytoplasm may lead to a better understanding of the processes governing actin dynamics within the cell and disease spread.
The mRNA expression of the cytokines interleukin (IL)-2, IL-4 and interferon (IFN)-γ and the production of IFN-γ was examined in mesenteric lymph node cells (MLNC) and CD4⁺ enriched cells populations from resistant and susceptible line lambs by use of reverse transcriptase-polymerase chain reactions (RT-PCR) and ELISA. MLNC obtained from lambs that were previously infected with a mixed gastro-intestinal nematode population were stimulated with Con A or Trichostrongylus colubriformis specific antigen. Four weeks after infection MLNC from both line lambs expressed high levels of mRNA coding for IL-2, IL-4 and IFN-γ. MLNC from resistant line lambs when stimulated with antigen for one day had higher mRNA expression of IL-2 and IFN-γ and after three days of culture had higher levels of IL-4 mRNA than MLNC from susceptible line lambs. Antigen stimulated MLNC of resistant line lambs had a higher IFN-γ production than those from susceptible line lambs. These results indicate that in resistant line lambs the secondary in vitro responses of cytokine mRNA and IFN-γ production to nematode specific antigen differs from those of susceptible line lambs.
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