• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jcmPermissionsJournals.ASM.orgJournalJCM ArticleJournal InfoAuthorsReviewers
J Clin Microbiol. Jun 1990; 28(6): 1385–1394.
PMCID: PMC267937

Growth and metabolism of Pneumocystis carinii in axenic culture.


Recent analyses of rRNA have suggested a taxonomic relationship of Pneumocystis carinii with fungi. We investigated various liquid and solid media without feeder cells for the ability to support growth of P. carinii. Organisms were obtained from the lungs of immunosuppressed rats and separated from host cells by treatment with hypotonic solutions and a series of microfiltrations. The growth of P. carinii was quantitated by staining nuclei with Diff-Quik. Optimal replication was observed in media with neopeptone and N-acetylglucosamine at low pH. Increases of at least 8- to 10-fold occurred during the first 24 to 72 h. P. carinii replication was influenced by pH, temperature, inoculum size, and anti-P. carinii drugs. The viability of organisms was confirmed by incorporation of radiolabeled uracil, methionine, and N-acetylglucosamine. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of trichloroacetic acid-precipitable compounds showed that [35S]methionine was incorporated into specific P. carinii proteins. Thus, P. carinii can metabolize and replicate in the absence of mammalian cells. This system should have applications for in vitro drug screening and other molecular and metabolic analyses.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • ANDREASEN AA, STIER TJB. Anaerobic nutrition of Saccharomyces cerevisiae. I. Ergosterol requirement for growth in a defined medium. J Cell Physiol. 1953 Feb;41(1):23–36. [PubMed]
  • ANDREASEN AA, STIER TJ. Anaerobic nutrition of Saccharomyces cerevisiae. II. Unsaturated fatty acid requirement for growth in a defined medium. J Cell Physiol. 1954 Jun;43(3):271–281. [PubMed]
  • Bartlett MS, Verbanac PA, Smith JW. Cultivation of Pneumocystis carinii with WI-38 cells. J Clin Microbiol. 1979 Dec;10(6):796–799. [PMC free article] [PubMed]
  • Barton EG, Jr, Campbell WG., Jr Further observations on the ultrastructure of pneumocystis. Arch Pathol. 1967 Jun;83(6):527–534. [PubMed]
  • Barton EG, Jr, Campbell WG., Jr Pneumocystis carinii in lungs of rats treated with cortisone acetate. Ultrastructural observations relating to the life cycle. Am J Pathol. 1969 Feb;54(2):209–236. [PMC free article] [PubMed]
  • Cushion MT. In vitro studies of Pneumocystis carinii. J Protozool. 1989 Jan-Feb;36(1):45–52. [PubMed]
  • Cushion MT, DeStefano JA, Walzer PD. Pneumocystis carinii: surface reactive carbohydrates detected by lectin probes. Exp Parasitol. 1988 Dec;67(2):137–147. [PubMed]
  • Cushion MT, Ruffolo JJ, Linke MJ, Walzer PD. Pneumocystis carinii: growth variables and estimates in the A549 and WI-38 VA13 human cell lines. Exp Parasitol. 1985 Aug;60(1):43–54. [PubMed]
  • Cushion MT, Stanforth D, Linke MJ, Walzer PD. Method of testing the susceptibility of Pneumocystis carinii to antimicrobial agents in vitro. Antimicrob Agents Chemother. 1985 Dec;28(6):796–801. [PMC free article] [PubMed]
  • Cushion MT, Walzer PD. Growth and serial passage of Pneumocystis carinii in the A549 cell line. Infect Immun. 1984 May;44(2):245–251. [PMC free article] [PubMed]
  • Desjardins RE, Canfield CJ, Haynes JD, Chulay JD. Quantitative assessment of antimalarial activity in vitro by a semiautomated microdilution technique. Antimicrob Agents Chemother. 1979 Dec;16(6):710–718. [PMC free article] [PubMed]
  • Edman JC, Kovacs JA, Masur H, Santi DV, Elwood HJ, Sogin ML. Ribosomal RNA sequence shows Pneumocystis carinii to be a member of the fungi. Nature. 1988 Aug 11;334(6182):519–522. [PubMed]
  • Frevert J, Ballou CE. Saccharomyces cerevisiae structural cell wall mannoprotein. Biochemistry. 1985 Jan 29;24(3):753–759. [PubMed]
  • Gigliotti F, Stokes DC, Cheatham AB, Davis DS, Hughes WT. Development of murine monoclonal antibodies to Pneumocystis carinii. J Infect Dis. 1986 Aug;154(2):315–322. [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]
  • Jackson PR, Pappas MG, Hansen BD. Fluorogenic substrate detection of viable intracellular and extracellular pathogenic protozoa. Science. 1985 Jan 25;227(4685):435–438. [PubMed]
  • Jones KH, Senft JA. An improved method to determine cell viability by simultaneous staining with fluorescein diacetate-propidium iodide. J Histochem Cytochem. 1985 Jan;33(1):77–79. [PubMed]
  • Kaneshiro ES, Cushion MT, Walzer PD, Jayasimhulu K. Analyses of Pneumocystis fatty acids. J Protozool. 1989 Jan-Feb;36(1):69S–72S. [PubMed]
  • Kovacs JA, Allegra CJ, Beaver J, Boarman D, Lewis M, Parrillo JE, Chabner B, Masur H. Characterization of de novo folate synthesis in Pneumocystis carinii and Toxoplasma gondii: potential for screening therapeutic agents. J Infect Dis. 1989 Aug;160(2):312–320. [PubMed]
  • 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]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Lewis TL, Keesler GA, Fenner GP, Parks LW. Pleiotropic mutations in Saccharomyces cerevisiae affecting sterol uptake and metabolism. Yeast. 1988 Jun;4(2):93–106. [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]
  • Matsumoto Y, Yoshida Y. Sporogony in Pneumocystis carinii: synaptonemal complexes and meiotic nuclear divisions observed in precysts. J Protozool. 1984 Aug;31(3):420–428. [PubMed]
  • Niimi M, Tokunaga M, Nakayama H. Regulation of mannitol catabolism in Candida albicans: evidence for cyclic AMP-independent glucose effect. J Med Vet Mycol. 1986 Jun;24(3):211–217. [PubMed]
  • Pesanti EL, Cox C. Metabolic and synthetic activities of Pneumocystis carinii in vitro. Infect Immun. 1981 Dec;34(3):908–914. [PMC free article] [PubMed]
  • Pifer LL, Woods D, Hughes WT. Propagation of Pneumocystis carinii in Vero cell culture. Infect Immun. 1978 Apr;20(1):66–68. [PMC free article] [PubMed]
  • Queener SF, Bartlett MS, Jay MA, Durkin MM, Smith JW. Activity of lipid-soluble inhibitors of dihydrofolate reductase against Pneumocystis carinii in culture and in a rat model of infection. Antimicrob Agents Chemother. 1987 Sep;31(9):1323–1327. [PMC free article] [PubMed]
  • Sprague GF, Jr, Blair LC, Thorner J. Cell interactions and regulation of cell type in the yeast Saccharomyces cerevisiae. Annu Rev Microbiol. 1983;37:623–660. [PubMed]
  • Stringer SL, Stringer JR, Blase MA, Walzer PD, Cushion MT. Pneumocystis carinii: sequence from ribosomal RNA implies a close relationship with fungi. Exp Parasitol. 1989 May;68(4):450–461. [PubMed]
  • Tegoshi T, Yoshida Y. New system of in vitro cultivation of Pneumocystis carinii without feeder cells. J Protozool. 1989 Jan-Feb;36(1):29S–31S. [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]
  • Vavra J, Kucera K. Pneumocystis carinii delanoë, its ultrastructure and ultrastructural affinities. J Protozool. 1970 Aug;17(3):463–483. [PubMed]
  • Vossen ME, Beckers PJ, Meuwissen JH, Stadhouders AM. Developmental biology of Pneumocystis carinii, and alternative view on the life cycle of the parasite. Z Parasitenkd. 1978 Apr 20;55(2):101–118. [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]
  • Walzer PD, Perl DP, Krogstad DJ, Rawson PG, Schultz MG. Pneumocystis carinii pneumonia in the United States. Epidemiologic, diagnostic, and clinical features. Ann Intern Med. 1974 Jan;80(1):83–93. [PubMed]
  • Wright AP, Bruns M, Hartley BS. Extraction and rapid inactivation of proteins from Saccharomyces cerevisiae by trichloroacetic acid precipitation. Yeast. 1989 Jan-Feb;5(1):51–53. [PubMed]
  • Yoshikawa H, Morioka H, Yoshida Y. Freeze-fracture localization of filipin-sterol complexes in plasma- and cyto-membranes of Pneumocystis carinii. J Protozool. 1987 May;34(2):131–137. [PubMed]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...