A novel lipoarabinomannan from the equine pathogen Rhodococcus equi. Structure and effect on macrophage cytokine production.
Rhodococcus equi is a major cause of foal morbidity and mortality. We have investigated the presence of lipoglycan in this organism as closely related bacteria, notably Mycobacterium tuberculosis, produce lipoarabinomannans (LAM) that may play multiple roles as virulence determinants. The lipoglycan was structurally characterized by gas chromatography-mass spectrometry following permethylation, capillary electrophoresis after chemical degradation, and (1)H and (31)P and two-dimensional heteronuclear nuclear magnetic resonance studies. Key structural features of the lipoglycan are a linear alpha-1,6-mannan with side chains containing one 2-linked alpha-d-Manp residue. This polysaccharidic backbone is linked to a phosphatidylinositol mannosyl anchor. In contrast to mycobacterial LAM, there are no extensive arabinan domains but single terminal alpha-d-Araf residue capping the 2-linked alpha-d-Manp. The lipoglycan binds concanavalin A and mannose-binding protein consistent with the presence of t-alpha-d-Manp residues. We studied the ability of the lipoglycans to induce cytokines from equine macrophages, in comparison to whole cells of R. equi. These data revealed patterns of cytokine mRNA induction that suggest that the lipoglycan is involved in much of the early macrophage cytokine response to R. equi infection. These studies identify a novel LAM variant that may contribute to the pathogenesis of disease caused by R. equi.
Garton
Natalie J
NJ
Institute of Pharmacy, Chemistry and Biomedical Sciences, the University of Sunderland, Sunderland SR2 3SD, United Kingdom.
Gilleron
Martine
M
Brando
Thérèse
T
Dan
Han-Hong
HH
Giguère
Steeve
S
Puzo
Germain
G
Prescott
John F
JF
Sutcliffe
Iain C
IC
D000818:Animals; D000942:Antigens, Bacterial / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D001483:Base Sequence; D002240:Carbohydrate Sequence; D016207:Cytokines / Q000276:immunology*; D017931:DNA Primers; D005227:Fatty Acids / Q000032:analysis; D005993:Glycerolphosphate Dehydrogenase / Q000235:genetics; D006736:Horses; D008070:Lipopolysaccharides / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D008264:Macrophages / Q000187:drug effects / Q000276:immunology*; D008745:Methylation; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000032:analysis; D009844:Oligosaccharides / Q000737:chemistry; D016929:Rhodococcus equi / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000942:Antigens, Bacterial; D016207:Cytokines; D017931:DNA Primers; D005227:Fatty Acids; D008070:Lipopolysaccharides; D009005:Monosaccharides; D009844:Oligosaccharides; C050016:lipoarabinomannan; D005993:Glycerolphosphate Dehydrogenase