Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1992 Dec;30(12):3263–3267. doi: 10.1128/jcm.30.12.3263-3267.1992

Purification of Pneumocystis carinii trophozoites and identification of their circulating antigens.

T Ishimaru 1, N Shimono 1, Y Sawae 1, Y Niho 1
PMCID: PMC270645  PMID: 1280653

Abstract

We have developed a new method to isolate Pneumocystis carinii trophozoites from experimental rat bronchoalveolar lavage specimens by using a Percoll discontinuous gradient and have identified the circulating antigens in experimental rat P. carinii pneumonia. The antigenic components of the trophozoites were compared with those of cysts by immunoblotting. A major immunoreactive band of 90,000 Da and some others of lower molecular mass were found in trophozoites. On the other hand, bands of 110,000, 50,000, and 45,000 Da were observed in cysts. The band of 50,000 Da was not identified when antitrophozoite rabbit serum preabsorbed with P. carinii-infected rat serum was used for immunoblotting. These results suggest that the molecule of 50,000 Da is the major circulating antigen in P. carinii-infected rats.

Full text

PDF
3263

Images in this article

3264Image
on p.3264
3265Image
on p.3265
3265Image
on p.3265
3266Image
on p.3266
3266Image
on p.3266

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Blumenfeld W., Griffiss J. M. In vitro differentiation of human-derived Pneumocystis carinii. J Clin Microbiol. 1989 Mar;27(3):480–485. doi: 10.1128/jcm.27.3.480-485.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Graves D. C. Immunological studies of Pneumocystis carinii. J Protozool. 1989 Jan-Feb;36(1):60–69. doi: 10.1111/j.1550-7408.1989.tb02700.x. [DOI] [PubMed] [Google Scholar]
  3. Graves D. C., McNabb S. J., Worley M. A., Downs T. D., Ivey M. H. Analyses of rat Pneumocystis carinii antigens recognized by human and rat antibodies by using western immunoblotting. Infect Immun. 1986 Oct;54(1):96–103. doi: 10.1128/iai.54.1.96-103.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Jarowenko M., Pifer L., Kerman R., Kahan B. D. Serologic methods for the early diagnosis of Pneumocystis carinii infection in renal allograft recipients. Transplantation. 1986 Apr;41(4):436–442. doi: 10.1097/00007890-198604000-00005. [DOI] [PubMed] [Google Scholar]
  5. Kim H. K., Hughes W. T., Feldman S. Studies of morphology and immunofluorescence of Pneumocystis carinii. Proc Soc Exp Biol Med. 1972 Oct;141(1):304–309. doi: 10.3181/00379727-141-36764. [DOI] [PubMed] [Google Scholar]
  6. Kovacs J. A., Halpern J. L., Lundgren B., Swan J. C., Parrillo J. E., 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. doi: 10.1093/infdis/159.1.60. [DOI] [PubMed] [Google Scholar]
  7. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  8. Linke M. J., Cushion M. T., Walzer P. D. Properties of the major antigens of rat and human Pneumocystis carinii. Infect Immun. 1989 May;57(5):1547–1555. doi: 10.1128/iai.57.5.1547-1555.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Maddison S. E., Hayes G. V., Ivey M. H., Tsang V. C., Slemenda S. B., Norman L. G. Fractionation of Pneumocystis carinii antigens used in an enzyme-linked immunosorbent assay for antibodies and in the production of antiserum for detecting Pneumocystis carinii antigenemia. J Clin Microbiol. 1982 Jun;15(6):1029–1035. doi: 10.1128/jcm.15.6.1029-1035.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Peglow S. L., Smulian A. G., Linke M. J., Pogue C. L., Nurre S., Crisler J., Phair J., Gold J. W., Armstrong D., Walzer P. D. Serologic responses to Pneumocystis carinii antigens in health and disease. J Infect Dis. 1990 Feb;161(2):296–306. doi: 10.1093/infdis/161.2.296. [DOI] [PubMed] [Google Scholar]
  11. Pifer L. W., Wolf B. L., Weems J. J., Jr, Woods D. R., Edwards C. C., Joyner R. E. Pneumocystis carinii antigenemia in acquired immunodeficiency syndrome. J Clin Microbiol. 1988 Jul;26(7):1357–1361. doi: 10.1128/jcm.26.7.1357-1361.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Tanabe K., Fuchimoto M., Egawa K., Nakamura Y. Use of Pneumocystis carinii genomic DNA clones for DNA hybridization analysis of infected human lungs. J Infect Dis. 1988 Mar;157(3):593–596. doi: 10.1093/infdis/157.3.593. [DOI] [PubMed] [Google Scholar]
  13. 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. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Wakefield A. E., Pixley F. J., Banerji S., Sinclair K., Miller R. F., Moxon E. R., Hopkin J. M. Detection of Pneumocystis carinii with DNA amplification. Lancet. 1990 Aug 25;336(8713):451–453. doi: 10.1016/0140-6736(90)92008-6. [DOI] [PubMed] [Google Scholar]
  15. Walzer P. D. Diagnosis of Pneumocystis carinii pneumonia. J Infect Dis. 1988 Apr;157(4):629–632. doi: 10.1093/infdis/157.4.629. [DOI] [PubMed] [Google Scholar]
  16. Walzer P. D., Linke M. J. A comparison of the antigenic characteristics of rat and human Pneumocystis carinii by immunoblotting. J Immunol. 1987 Apr 1;138(7):2257–2265. [PubMed] [Google Scholar]
  17. Walzer P. D., Stanforth D., Linke M. J., Cushion M. T. Pneumocystis carinii: immunoblotting and immunofluorescent analyses of serum antibodies during experimental rat infection and recovery. Exp Parasitol. 1987 Jun;63(3):319–328. doi: 10.1016/0014-4894(87)90179-2. [DOI] [PubMed] [Google Scholar]
  18. Watanabe J., Tanabe K., Shimada K. Separation of trophozoites of Pneumocystis carinii from lung of the inoculated nude rat. Jpn J Exp Med. 1987 Oct;57(5):295–297. [PubMed] [Google Scholar]

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

RESOURCES