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J Clin Invest. Jan 1991; 87(1): 163–170.
PMCID: PMC295016

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.

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  • Kovacs JA, Halpern JL, Swan JC, Moss J, Parrillo JE, Masur H. Identification of antigens and antibodies specific for Pneumocystis carinii. J Immunol. 1988 Mar 15;140(6):2023–2031. [PubMed]
  • Kovacs JA, Halpern JL, Lundgren B, Swan JC, Parrillo JE, Masur H. Monoclonal antibodies to Pneumocystis carinii: identification of specific antigens and characterization of antigenic differences between rat and human isolates. J Infect Dis. 1989 Jan;159(1):60–70. [PubMed]
  • Walzer PD, Linke MJ. A comparison of the antigenic characteristics of rat and human Pneumocystis carinii by immunoblotting. J Immunol. 1987 Apr 1;138(7):2257–2265. [PubMed]
  • Graves DC, McNabb SJ, Ivey MH, Worley MA. Development and characterization of monoclonal antibodies to Pneumocystis carinii. Infect Immun. 1986 Jan;51(1):125–133. [PMC free article] [PubMed]
  • Graves DC, McNabb SJ, Worley MA, Downs TD, Ivey MH. Analyses of rat Pneumocystis carinii antigens recognized by human and rat antibodies by using western immunoblotting. Infect Immun. 1986 Oct;54(1):96–103. [PMC free article] [PubMed]
  • Radding JA, Armstrong MY, Ullu E, Richards FF. Identification and isolation of a major cell surface glycoprotein of Pneumocystis carinii. Infect Immun. 1989 Jul;57(7):2149–2157. [PMC free article] [PubMed]
  • Gigliotti F, Ballou LR, Hughes WT, Mosley BD. Purification and initial characterization of a ferret Pneumocystis carinii surface antigen. J Infect Dis. 1988 Oct;158(4):848–854. [PubMed]
  • Linke MJ, Cushion MT, Walzer PD. Properties of the major antigens of rat and human Pneumocystis carinii. Infect Immun. 1989 May;57(5):1547–1555. [PMC free article] [PubMed]
  • Pesanti EL, Shanley JD. Glycoproteins of Pneumocystis carinii: characterization by electrophoresis and microscopy. J Infect Dis. 1988 Dec;158(6):1353–1359. [PubMed]
  • Tanabe K, Takasaki S, Watanabe J, Kobata A, Egawa K, Nakamura Y. Glycoproteins composed of major surface immunodeterminants of Pneumocystis carinii. Infect Immun. 1989 May;57(5):1363–1368. [PMC free article] [PubMed]
  • Masur H, Jones TC. The interaction in vitro of Pneumocystis carinii with macrophages and L-cells. J Exp Med. 1978 Jan 1;147(1):157–170. [PMC free article] [PubMed]
  • Kitamura K, Yamamoto Y. Purification and properties of an enzyme, zymolyase, which lyses viable yeast cells. Arch Biochem Biophys. 1972 Nov;153(1):403–406. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PMC free article] [PubMed]
  • Maley F, Trimble RB, Tarentino AL, Plummer TH., Jr Characterization of glycoproteins and their associated oligosaccharides through the use of endoglycosidases. Anal Biochem. 1989 Aug 1;180(2):195–204. [PubMed]
  • Trimble RB, Maley F. Optimizing hydrolysis of N-linked high-mannose oligosaccharides by endo-beta-N-acetylglucosaminidase H. Anal Biochem. 1984 Sep;141(2):515–522. [PubMed]
  • Clegg JC. Glycoprotein detection in nitrocellulose transfers of electrophoretically separated protein mixtures using concanavalin A and peroxidase: application to arenavirus and flavivirus proteins. Anal Biochem. 1982 Dec;127(2):389–394. [PubMed]
  • Staros JV. N-hydroxysulfosuccinimide active esters: bis(N-hydroxysulfosuccinimide) esters of two dicarboxylic acids are hydrophilic, membrane-impermeant, protein cross-linkers. Biochemistry. 1982 Aug 17;21(17):3950–3955. [PubMed]
  • Gigliotti F, Hughes WT. Passive immunoprophylaxis with specific monoclonal antibody confers partial protection against Pneumocystis carinii pneumonitis in animal models. J Clin Invest. 1988 Jun;81(6):1666–1668. [PMC free article] [PubMed]
  • Yoshikawa H, Tegoshi T, Yoshida Y. Detection of surface carbohydrates on Pneumocystis carinii by fluorescein-conjugated lectins. Parasitol Res. 1987;74(1):43–49. [PubMed]
  • Cabib E, Roberts R, Bowers B. Synthesis of the yeast cell wall and its regulation. Annu Rev Biochem. 1982;51:763–793. [PubMed]
  • Norman L, Kagan IG. Some observations on the serology of Pneumocystis carinii infections in the United States. Infect Immun. 1973 Sep;8(3):317–321. [PMC free article] [PubMed]
  • Meuwissen JH, Tauber I, Leeuwenberg AD, Beckers PJ, Sieben M. Parasitologic and serologic observations of infection with Pneumocystis in humans. J Infect Dis. 1977 Jul;136(1):43–49. [PubMed]
  • Tanabe K, Furuta T, Ueda K, Tanaka H, Shimada K. Serological observations of Pneumocystis carinii infection in humans. J Clin Microbiol. 1985 Dec;22(6):1058–1060. [PMC free article] [PubMed]
  • Pifer LL, Hughes WT, Stagno S, Woods D. Pneumocystis carinii infection: evidence for high prevalence in normal and immunosuppressed children. Pediatrics. 1978 Jan;61(1):35–41. [PubMed]
  • Maddison SE, Hayes GV, Slemenda SB, Norman LG, Ivey MH. Detection of specific antibody by enzyme-linked immunosorbent assay and antigenemia by counterimmunoelectrophoresis in humans infected with Pneumocystis carinii. J Clin Microbiol. 1982 Jun;15(6):1036–1043. [PMC free article] [PubMed]
  • Peglow SL, Smulian AG, Linke MJ, Pogue CL, Nurre S, Crisler J, Phair J, Gold JW, Armstrong D, Walzer PD. Serologic responses to Pneumocystis carinii antigens in health and disease. J Infect Dis. 1990 Feb;161(2):296–306. [PubMed]
  • Herrod HG, Valenski WR, Woods DR, Pifer LL. The in vitro response of human lymphocytes to Pneumocystis carinii antigen. J Immunol. 1981 Jan;126(1):59–61. [PubMed]
  • Hagler DN, Deepe GS, Pogue CL, Walzer PD. Blastogenic responses to Pneumocystis carinii among patients with human immunodeficiency (HIV) infection. Clin Exp Immunol. 1988 Oct;74(1):7–13. [PMC free article] [PubMed]
  • Hofmann B, Nielsen PB, Odum N, Gerstoft J, Platz P, Ryder LP, Poulsen AG, Mathiesen L, Dickmeiss E, Norrild B, et al. Humoral and cellular responses to Pneumocystis carinii, CMV, and herpes simplex in patients with AIDS and in controls. Scand J Infect Dis. 1988;20(4):389–394. [PubMed]
  • Pottratz ST, Martin WJ., 2nd Role of fibronectin in Pneumocystis carinii attachment to cultured lung cells. J Clin Invest. 1990 Feb;85(2):351–356. [PMC free article] [PubMed]
  • Stahl P, Schlesinger PH, Sigardson E, Rodman JS, Lee YC. Receptor-mediated pinocytosis of mannose glycoconjugates by macrophages: characterization and evidence for receptor recycling. Cell. 1980 Jan;19(1):207–215. [PubMed]
  • Stahl PD, Rodman JS, Miller MJ, Schlesinger PH. Evidence for receptor-mediated binding of glycoproteins, glycoconjugates, and lysosomal glycosidases by alveolar macrophages. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1399–1403. [PMC free article] [PubMed]

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