The phosphatidyl-myo-inositol anchor of the lipoarabinomannans from Mycobacterium bovis bacillus Calmette Guérin. Heterogeneity, structure, and role in the regulation of cytokine secretion.
Lipoarabinomannans are major mycobacterial antigens capable of modulating the host immune response; however, the molecular basis underlying the diversity of their immunological properties remain an open question. In this study a new extraction and purification approach was successfully applied to isolate ManLAMs (lipoarabinomannans with mannosyl extensions) from bacillus Calmette Guérin leading to the obtention of two types of ManLAMs namely parietal and cellular. Structurally, they were found to differ by the percentage of mannooligosaccharide caps, 76 and 48%, respectively, and also, thanks to a new analytical method, by the structure of the phosphatidyl-myo-inositol anchor lipid moiety. A novel fatty acid in the mycobacterium genus assigned to a 12-O-(methoxypropanoyl)-12-hydroxystearic acid was the only fatty acid esterifying C-1 of the glycerol residue of the parietal ManLAMs, while the phosphatidyl unit of the cellular ManLAMs showed a large heterogeneity due to a combination of palmitic and tuberculostearic acid. Finally, parietal and cellular ManLAMs were found to differentially affect interleukin-8 and tumor necrosis factor-alpha secretion from human dendritic cells. We show that parietal but not cellular ManLAMs were able to stimulate tumor necrosis factor-alpha secretion from dendritic cells. From these studies we propose that the 1-[12-O-(methoxypropanoyl)-12-hydroxystearoyl]-sn-glycerol part is the major cytokine-regulating component of the ManLAMs. It seems likely that modification of the ManLAM lipid part, which may occur in hostile environments, could regulate macrophagic mycobacterial survival by altering cytokine stimulation.
Nigou
J
J
Institut de Pharmacologie et de Biologie Structurale du Centre National de la Recherche Scientifique, 118 route de Narbonne, 31062 Toulouse Cedex, France.
Gilleron
M
M
Cahuzac
B
B
Bounéry
J D
JD
Herold
M
M
Thurnher
M
M
Puzo
G
G
D000942:Antigens, Bacterial / Q000737:chemistry* / Q000276:immunology; D001422:Bacterial Adhesion; D002240:Carbohydrate Sequence; D002478:Cells, Cultured; D002849:Chromatography, Gas; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology; D008401:Gas Chromatography-Mass Spectrometry; D017261:Glycosylphosphatidylinositols / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D037181:Lectins, C-Type; D008070:Lipopolysaccharides / Q000737:chemistry* / Q000276:immunology; D009682:Magnetic Resonance Spectroscopy; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D008969:Molecular Sequence Data; D009163:Mycobacterium bovis / Q000737:chemistry* / Q000276:immunology; D019308:Palmitic Acid / Q000032:analysis; D010586:Phagocytes / Q000276:immunology / Q000382:microbiology; D011956:Receptors, Cell Surface; D013229:Stearic Acids / Q000032:analysis; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
C108079:12-O-(methoxypropanoyl)-12-hydroxystearic acid; D000942:Antigens, Bacterial; D017261:Glycosylphosphatidylinositols; D016209:Interleukin-8; D037181:Lectins, C-Type; D008070:Lipopolysaccharides; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D011956:Receptors, Cell Surface; D013229:Stearic Acids; D014409:Tumor Necrosis Factor-alpha; C050016:lipoarabinomannan; D019308:Palmitic Acid; C026082:tuberculostearic acid