Purification and characterization of a major human Pneumocystis carinii surface antigen.
Previous studies of Pneumocystis carinii have identified the major surface antigen of rat and human isolates as proteins of 116,000 and 95,000 mol wt, respectively, that are antigenically not identical. In this study both rat and human P. carinii proteins were purified by solubilization with zymolyase followed by molecular sieve and ion exchange chromatography. The native proteins had an apparent mol wt of 290,000 or greater, based on molecular sieve studies as well as cross-linking studies. Both proteins were glycoproteins; treatment with endoglycosidase H resulted in a 9% decrease in mol wt. The carbohydrate composition of the rat P. carinii glycoprotein was distinct from the human isolate; glucose, mannose, galactose, and glucosamine occurred in approximately equimolar ratios in the human P. carinii protein, whereas glucose and mannose were the predominant sugars of the rat P. carinii protein. To evaluate humoral immune responses to the human P. carinii protein, an enzyme-linked immunosorbent assay using purified protein was developed. Some, but not all, patients who subsequently developed P. carinii pneumonia demonstrated a serum antibody response to the surface antigen. Nearly all subjects without a history of P. carinii pneumonia had no detectable antibodies. Purified P. carinii proteins will greatly facilitate the investigation of host-P. carinii interactions.
Lundgren
B
B
Critical Care Medicine Department, National Institutes of Health, Bethesda, Maryland 20892.
Lipschik
G Y
GY
Kovacs
J A
JA
D000818:Animals; D000946:Antigens, Fungal / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D000954:Antigens, Surface / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay; D008358:Mannose / Q000032:analysis; D008970:Molecular Weight; D011010:Pneumocystis / Q000276:immunology*; D011020:Pneumonia, Pneumocystis / Q000276:immunology; D051381:Rats; D011919:Rats, Inbred Strains
D000946:Antigens, Fungal; D000954:Antigens, Surface; D008358:Mannose