Purification of the 115-kilodalton exoantigen of Cryptococcus neoformans and its recognition by immune sera.
A 115-kDa exoantigen produced by Cryptococcus neoformans recognized by the previously described murine monoclonal antibody 7C9 has been purified from culture filtrate by a combination of membrane ultrafiltration, isoelectric focusing, and preparative gel electrophoresis. It is produced in late-log-phase cultures and is present in greater amounts in cultures grown at 25 degrees C than in those grown at 37 degrees C. Recognition of the antigen by 7C9 on immunoblots is abolished by the proteolytic enzymes papain and trypsin. The antigen is a glycoprotein bearing N-linked oligosaccharides, of which mannose is an important constituent. It does not appear to have proteolytic activity and is acidic, with a pI of 3 to 3.2. Its relationship to previously described C. neoformans mannoprotein is unclear since 7C9 shows only very weak cross-reactivity with a purified sample of the latter. Sera from patients infected with C. neoformans exhibited strong recognition of the glycoprotein as shown by immunoenzyme development of Western immunoblots, indicating its possible significance as a marker of disease.
Hamilton
A J
AJ
Dermatology Unit, St. Johns Institute of Dermatology, Guys Hospital, London, United Kingdom.
Goodley
J
J
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial / Q000737:chemistry / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D015415:Biomarkers; D003453:Cryptococcosis / Q000175:diagnosis; D003455:Cryptococcus neoformans / Q000276:immunology*; D005260:Female; D006023:Glycoproteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D006801:Humans; D008297:Male; D008358:Mannose / Q000032:analysis; D008970:Molecular Weight
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D015415:Biomarkers; D006023:Glycoproteins; D008358:Mannose