Purification and characterization of the serotype-specific polysaccharide antigen of Trichosporon cutaneum serotype II: a disease-related antigen of Japanese summer-type hypersensitivity pneumonitis.
Summer-type hypersensitivity pneumonitis (SHP) is a unique type of hypersensitivity pneumonitis and the most prevalent in Japan. Our previous study clarified that the causative agent of the disease is Trichosporon cutaneum, and that the patients with SHP have high titres of antibodies against the serotype-specific antigen of polysaccharide nature which exist in the high molecular weight fraction of the culture supernatant of the yeast. In this study, we purified the serotype-specific antigen of serotype II T. cutaneum by gel filtration and affinity chromatography using a monoclonal antibody, D-8, specific for a high molecular weight antigen of serotype II T. cutaneum, and elucidated the structure of the antigen. This affinity-purified antigen was shown to be an essentially acidic polysaccharide comprising mannose, xylose, and glucuronic acid (6:44:4.7). Chemical analysis showed that this polysaccharide antigen contains a (1-3)-linked mannan backbone attached with short side chains of (1-4)-linked mannose and a small proportion of (1-2)-linked xylose residues by substituting the 2- or 4-positions of the (1-3)-linked mannose residues of the main chain. Approximately one-fifth of the side chains were terminated with glucuronic acid residues. The antigenic epitope of the serotype-specific antigen was shown to involve the terminal glucoronic acid residues as revealed by immunodiffusion test and sandwich enzyme-linked immunosorbent assay using monoclonal antibody D-8.
Mizobe
T
T
Department of Immunology, Kumamoto University School of Medicine, Japan.
Ando
M
M
Yamasaki
H
H
Onoue
K
K
Misaki
A
A
D000542:Alveolitis, Extrinsic Allergic / Q000209:etiology*; D000911:Antibodies, Monoclonal; D000946:Antigens, Fungal / Q000009:adverse effects / Q000737:chemistry / Q000302:isolation & purification*; D002240:Carbohydrate Sequence; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005965:Glucuronates / Q000032:analysis; D020723:Glucuronic Acid; D006801:Humans; D007564:Japan; D008358:Mannose / Q000032:analysis; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000009:adverse effects / Q000737:chemistry / Q000302:isolation & purification*; D012703:Serotyping; D014250:Trichosporon / Q000145:classification / Q000276:immunology*; D014994:Xylose / Q000032:analysis
D000911:Antibodies, Monoclonal; D000946:Antigens, Fungal; D000939:Epitopes; D005965:Glucuronates; D011134:Polysaccharides; D020723:Glucuronic Acid; D014994:Xylose; D008358:Mannose