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1.
Experimental & Molecular Medicine ; : 538-538, 2011.
Article in English | WPRIM | ID: wpr-7974

ABSTRACT

No abstract available.

2.
Infection and Chemotherapy ; : 23-29, 2010.
Article in Korean | WPRIM | ID: wpr-225194

ABSTRACT

BACKGROUND: Legionella pneumophila is the causative agent of Legionnaires' disease, a severe form of pneumonia. After L. pneumophila is inhaled through contaminated aerosols, it is phagocytized by alveolar macrophages, multiplies in a specialized phagosome approximately 10 h postinfection, and eventually leads to the death of host cells. Currently available diagnostic tests for Legionella pneumonia have some limitations. This study was conducted to find diagnostic biomarkers for Legionella pneumonia using virulence gene expression profiling in a murine experimental model. MATERIALS AND METHODS: A/J mice were intranasally inoculated with L. pneumophila serogroup 1, and lungs were harvested 4, 8, 24, and 48 h postinfection. The strain grown in buffered yeast extract broth was used as reference samples. Cy-dye labeled cDNA samples were prepared with total RNA from lungs or broth culture, and hybridized on the oligo-microarray slide containing 2,895 genes of L. pneumophila serogroup 1. Virulence gene expression patterns were analyzed using a MIDAS software from TIGR (www.tigr.org). RESULTS: Among a total of 332 virulence genes examined, 17 genes including sidA, lepB, the genes related to flagella assembly (fliR and fliP), LPS lipid A biosynthesis, and the enhanced entry protein EnhA were up-regulated at all four time points. We further confirmed by quantitative real-time reverse transcription PCR that the expression of fliP gene was highly expressed in lung tissue as well as in bronchoalveolar lavage fluids from the mouse infected with L. pneumophila serogroup 1. CONCLUSIONS: Through gene expression analysis of L. pneumophila in a mouse model, several candidate biomarkers for diagnosing Legionnaires' disease could be identified.


Subject(s)
Animals , Mice , Aerosols , Biomarkers , Bronchoalveolar Lavage Fluid , Chimera , Diagnostic Tests, Routine , DNA, Complementary , Flagella , Gene Expression , Gene Expression Profiling , Legionella , Legionella pneumophila , Legionnaires' Disease , Lipid A , Lung , Macrophages, Alveolar , Oligonucleotide Array Sequence Analysis , Phagosomes , Pneumonia , Polymerase Chain Reaction , Reverse Transcription , RNA , Sprains and Strains , Yeasts
3.
Experimental & Molecular Medicine ; : 687-694, 2009.
Article in English | WPRIM | ID: wpr-71514

ABSTRACT

Legionella bacterium, an intracellular pathogen of mononuclear phagocytes, causes acute fatal pneumonia, especially in patients with impaired cellular immune responses. Until recently, however, the toll-like receptor (TLR) engagement of bacterial proteins derived from Legionella is uncertain. We previously showed that a 19-kDa highly conserved peptidoglycan-associated lipoprotein (PAL) of Legionella pneumophila induced the PAL-specific B cell and T cell responses in mice. In this study, we observed that the rPAL antigen of L. pneumophila, as an effector molecule, activated murine macrophages via TLR2 and produced proinflammatory cytokines such as IL-6 and TNF-alpha. In both BALB/c and TLR4-deficient C3H/HeJ mice, pretreatment of macrophages with anti-TLR2 mAb showed severely impaired cytokine production in response to the rPAL. In addition, in vitro the rPAL treatment increased the cell surface expression of CD40, CD80, CD86 and MHC I/II molecules. We further showed that the synthetic CpG-oligodeoxynucleotides (CpG ODN) coadministered with the rPAL enhanced IL-12 and IL-6 production and expression of CD40, CD80 and MHC II compared to the rPAL treatment alone. In conclusions, these results indicate that Legionella PAL might activate macrophages via a TLR2-dependent mechanism which thus induce cytokine production and expression of costimulatory and MHC molecules.


Subject(s)
Animals , Female , Mice , Antigens, CD/immunology , Bacterial Outer Membrane Proteins/pharmacology , Cells, Cultured , Histocompatibility Antigens Class II/immunology , Host-Pathogen Interactions , Interleukin-12/biosynthesis , Interleukin-6/biosynthesis , Legionella pneumophila/immunology , Legionnaires' Disease/immunology , Lipoproteins/pharmacology , Macrophage Activation/drug effects , Macrophages, Peritoneal/drug effects , Mice, Inbred BALB C , Mice, Inbred C3H , Mice, Inbred C57BL , Toll-Like Receptor 2/metabolism , Tumor Necrosis Factor-alpha/biosynthesis
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