Abstract
Tsukamurella pulmonis is an opportunistic actinomycetal pathogen associated with a variety of rarely diagnosed human infections. In clinical cases of infection, T. pulmonis usually accompanies other bacterial pathogens. Because of these mixed infections, a robust diagnostic assay is important. The bacteria cell surface polysaccharides are considered not only useful targets for diagnostics but also intriguing subjects for analysis of the interactions that regulate the host response in general. Here, the structure of the polysaccharide component of the T. pulmonis cell wall was established. Sugar and methylation analysis and 2D-NMR techniques revealed that its polysaccharide belongs to the class of arabinomannan composed of branched tetrasaccharide repeating units, with addition of linear →6)-α-D-Manp-(1→ mannan. Rabbit polyclonal sera against T. pulmonis and T. paurometabola bacterial cells revealed cross reactivity between their antigens. Tissue samples from mice infected with T. pulmonis revealed liver abscesses and pathologic granules located intracellularly when immunohistochemically stained with monoclonal antibodies raised against T. pulmonis polysaccharide. Ultrastructural studies revealed that these granules contain T. pulmonis cells. These observations indicate that T. pulmonis is a pathogenic species capable of spreading within the organism, presumably through the blood.
Highlights
Tsukamurella strains are opportunistic bacteria and are often accompanied by other pathogens
The objective of this study was to determine the structure of the polysaccharide antigen from the cell wall of T. pulmonis and generate antibodies against this polysaccharide, which will be useful for studying bacterial infection of host tissue using immunohistochemical and electron microscopic identification of the distribution of polysaccharide epitopes both on the bacteria and within tissues infected with this pathogen
Members of the Tsukamurella genus are known as opportunistic pathogens and are agents of lung cavitations and tuberculosis-like syndromes[33]
Summary
Tsukamurella strains are opportunistic bacteria and are often accompanied by other pathogens. Reclassification of species within the genus Tsukamurella has been proposed—T. spongiae should be reclassified as a later heterotypic synonym of T. pulmonis, T. carboxydivorans as a synonym of T. tyrosinosolvens, and T. sunchonensis as a later heterotypic synonym of T. pseudospumae. All these studies were based on phenotypic characteristics and on comparison of whole bacterial genomes[16]. The objective of this study was to determine the structure of the polysaccharide antigen from the cell wall of T. pulmonis and generate antibodies against this polysaccharide, which will be useful for studying bacterial infection of host tissue using immunohistochemical and electron microscopic identification of the distribution of polysaccharide epitopes both on the bacteria and within tissues infected with this pathogen
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