Antigenic properties of recombinant glycosylated and nonglycosylated Pneumocystis carinii glycoprotein A polypeptides expressed in baculovirus-infected insect cells.
Since a continuous culture system is not yet available for the opportunistic fungal pathogen Pneumocystis carinii, obtaining suitable amounts of purified P. carinii antigens free of mammalian-host lung contaminants is difficult. Hence, production of recombinant antigen possessing epitopes found in native P. carinii antigens is critical for immunological studies. We utilized the baculovirus expression vector system (BEVS) in insect cells to determine whether B-cell epitopes present in the protein core of a native P. carinii surface glycoprotein were conserved in the recombinant polypeptide, and to investigate its glycosylation by insect cells. B-cell epitopes were retained, but the insect cells appeared to hyperglycosylate the recombinant protein.
Haidaris
C G
CG
Department of Microbiology and Immunology, University of Rochester, School of Medicine and Dentistry, NY 14642, USA. haid@uhura.cc.rochester.edu
Fisher
D J
DJ
Gigliotti
F
F
Simpson-Haidaris
P J
PJ
D000818:Animals; D000911:Antibodies, Monoclonal; D000946:Antigens, Fungal / Q000276:immunology*; D016367:Baculoviridae / Q000235:genetics*; D015153:Blotting, Western; D002460:Cell Line; D000939:Epitopes / Q000096:biosynthesis; D005289:Ferrets / Q000382:microbiology; D005656:Fungal Proteins / Q000096:biosynthesis / Q000276:immunology* / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D006031:Glycosylation / Q000187:drug effects; D008168:Lung / Q000382:microbiology; D008297:Male; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000276:immunology* / Q000378:metabolism; D011010:Pneumocystis / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism; D018411:Spodoptera; D014415:Tunicamycin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000946:Antigens, Fungal; D000939:Epitopes; D005656:Fungal Proteins; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D014415:Tunicamycin; C077556:GPA protein, Pneumocystis carinii sp.