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ORIGINAL RESEARCH ARTICLE

Contribution of Fucose-Containing Capsules in Klebsiella pneumoniae to Bacterial Virulence in Mice

June Hsieh Wu^*,1, Albert M. Wu, Cheng Gie Tsai^*, Xin-Yu Chang^*, Shih-Feng Tsai and Ting-Shu Wu

^* Department of Microbiology and Immunology, College of Medicine, Chang Gung University, Tao Yuan 333, Taiwan; Glyco-Immunochemistry Research Laboratory, Institute of Molecular and Cellular Biology, Chang Gung University, Tao Yuan 333, Taiwan; Department of Molecular and Genomic Medicine, National Health Research Institute, Miaoli 350, Taiwan; and Division of Infectious Disease, Department of Internal Medicine, Chang Gung Memorial Hospital, Tao Yuan 333, Taiwan

To whom requests for reprints should be addressed at ¹ Department of Microbiology and Immunology, College of Medicine, Chang Gung University, 259 Wen Hua 1 Road, Kwei San, Tao Yuan 333 Taiwan. E-mail: jhwu{at}mail.cgu.edu.tw

Bacterium Klebsiella pneumoniae (KP) contains a prominent capsule. Clinical infections usually are associated with pneumonia or urinary tract infection (UTI). Emerging evidence implicates KP in severe liver abscess especially in diabetic patients. The goal of this study was to investigate the capsular polysaccharides from KP of liver abscess (hepatic-KP) and of UTI-KP. The composition of capsular polysaccharides was analyzed by capillary high-performance liquid chromatography (HPLC, Dionex system). The terminal sugars were assayed by binding ability to lectins. The results showed that the capsule of a hepatic KP (KpL1) from a diabetic patient contained fucose, while the capsule from UTI-KP (KpU1) did not. The absence of fucose was verified by the absence of detectable polymerase chain reaction (PCR) fragment for fucose synthesis genes, gmd and wcaG in KpU1. Mice infected with the KpL1 showed high fatality, whereas those infected with the KpU1 showed high survival rate. The KpL1 capsule was reactive to lectins AAA and AAL, which detect fucose, while the KpU1 capsule was reactive to lectin GNA, which detects mannose. Phagocytosis experiment in mouse peritoneal cavity indicated that the peritoneal macrophages could interact with KpU1, while rare association of KpL1 with macrophages was observed. This study revealed that different polysaccharides were displayed on the bacterial capsules of virulent KpL1 as compared with the less virulent KpU1. Interaction of KpU1 with mice peritoneal macrophages was more prominent than that of KpL1. The possession of fucose might contribute to KpL1 virulence by avoiding phagocytosis since fucose on bacteria had been implicated in immune evasion.

Key Words: bacterial capsule • diabetes mellitus • enzyme-linked immunosorbent assay • fucose • lectin binding assay • liver abscess