Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1996 Jan;64(1):290–297. doi: 10.1128/iai.64.1.290-297.1996

Antigenic differences associated with genetically distinct Pneumocystis carinii from rats.

J Vasquez 1, A G Smulian 1, M J Linke 1, M T Cushion 1
PMCID: PMC173758  PMID: 8557354

Abstract

Pneumocystis carinii is a family of organisms found in a wide variety of mammalian lungs. In immunocompromised hosts, the organisms are able to produce an oftentimes fatal pneumonia. The existence of distinct types of Pneumocystis populations is strongly supported by antigenic and genetic evidence. In the present study, we assessed the antigenic profiles of two genetically distinct Pneumocystis carinii populations, P. carinii f. sp. carinii and P. carinii f. sp. ratti, as well as two types of P. carinii f. sp. carinii defined by electrophoretic karyotyping (forms 1 and 2). The separated and blotted proteins of the organism preparations were probed with four monoclonal antibodies (MAbs) generated to the major surface glycoproteins of rat-derived P. carinii, one anti-human P. carinii MAb, and two polyclonal antisera made with rat-derived P. carinii as the immunogen. Differences in reactivities between the P. carinii f. sp. carinii and P. carinii f. sp. ratti preparations were detected with two of the MAbs, and both of the rat P. carinii polyclonal antisera in the 45- to 55-kDa molecular mass range, but not with the human P. carinii MAb. The reactivities of the 16 P. carinii f. sp. carinii preparations were the same with two exceptions. Two preparations of form 1 showed strong reactivity with the anti-MSG MAb RA-C11. The ratios of cyst forms to trophic forms evaluated by microscopy were not associated with any of the differences observed in the antigenic profiles. The antigenic differences between P. carinii f. sp. carinii and P. carinii f. sp. ratti are consistent with the distinction of these two populations made by molecular genetic techniques, while the two differences detected among the P. carinii f. sp. carinii preparations suggest the organism may be able to modulate antigenic epitopes. The use of immunoblotting to differentiate infecting organism populations and assess antigenic modulation holds promise for future epidemiologic studies.

Full Text

The Full Text of this article is available as a PDF (339.1 KB).

Selected References

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

  1. Boylan C. J., Current W. L. Improved rat model of Pneumocystis carinii pneumonia: induced laboratory infections in Pneumocystis-free animals. Infect Immun. 1992 Apr;60(4):1589–1597. doi: 10.1128/iai.60.4.1589-1597.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cushion M. T., Kaselis M., Stringer S. L., Stringer J. R. Genetic stability and diversity of Pneumocystis carinii infecting rat colonies. Infect Immun. 1993 Nov;61(11):4801–4813. doi: 10.1128/iai.61.11.4801-4813.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cushion M. T., Ruffolo J. J., Linke M. J., Walzer P. D. Pneumocystis carinii: growth variables and estimates in the A549 and WI-38 VA13 human cell lines. Exp Parasitol. 1985 Aug;60(1):43–54. doi: 10.1016/s0014-4894(85)80021-7. [DOI] [PubMed] [Google Scholar]
  4. Cushion M. T., Zhang J., Kaselis M., Giuntoli D., Stringer S. L., Stringer J. R. Evidence for two genetic variants of Pneumocystis carinii coinfecting laboratory rats. J Clin Microbiol. 1993 May;31(5):1217–1223. doi: 10.1128/jcm.31.5.1217-1223.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Edlind T. D., Bartlett M. S., Weinberg G. A., Prah G. N., Smith J. W. The beta-tubulin gene from rat and human isolates of Pneumocystis carinii. Mol Microbiol. 1992 Nov;6(22):3365–3373. doi: 10.1111/j.1365-2958.1992.tb02204.x. [DOI] [PubMed] [Google Scholar]
  6. Edman J. C., Kovacs J. A., Masur H., Santi D. V., Elwood H. J., Sogin M. L. Ribosomal RNA sequence shows Pneumocystis carinii to be a member of the fungi. Nature. 1988 Aug 11;334(6182):519–522. doi: 10.1038/334519a0. [DOI] [PubMed] [Google Scholar]
  7. Ezekowitz R. A., Williams D. J., Koziel H., Armstrong M. Y., Warner A., Richards F. F., Rose R. M. Uptake of Pneumocystis carinii mediated by the macrophage mannose receptor. Nature. 1991 May 9;351(6322):155–158. doi: 10.1038/351155a0. [DOI] [PubMed] [Google Scholar]
  8. Fisher D. J., Gigliotti F., Zauderer M., Harmsen A. G. Specific T-cell response to a Pneumocystis carinii surface glycoprotein (gp120) after immunization and natural infection. Infect Immun. 1991 Oct;59(10):3372–3376. doi: 10.1128/iai.59.10.3372-3376.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gigliotti F., Ballou L. R., Hughes W. T., Mosley B. D. Purification and initial characterization of a ferret Pneumocystis carinii surface antigen. J Infect Dis. 1988 Oct;158(4):848–854. doi: 10.1093/infdis/158.4.848. [DOI] [PubMed] [Google Scholar]
  10. Gigliotti F. Host species-specific antigenic variation of a mannosylated surface glycoprotein of Pneumocystis carinii. J Infect Dis. 1992 Feb;165(2):329–336. doi: 10.1093/infdis/165.2.329. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Hong S. T., Steele P. E., Cushion M. T., Walzer P. D., Stringer S. L., Stringer J. R. Pneumocystis carinii karyotypes. J Clin Microbiol. 1990 Aug;28(8):1785–1795. doi: 10.1128/jcm.28.8.1785-1795.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Keely S. P., Stringer J. R., Baughman R. P., Linke M. J., Walzer P. D., Smulian A. G. Genetic variation among Pneumocystis carinii hominis isolates in recurrent pneumocystosis. J Infect Dis. 1995 Aug;172(2):595–598. doi: 10.1093/infdis/172.2.595. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Kovacs J. A., Powell F., Edman J. C., Lundgren B., Martinez A., Drew B., Angus C. W. Multiple genes encode the major surface glycoprotein of Pneumocystis carinii. J Biol Chem. 1993 Mar 15;268(8):6034–6040. [PubMed] [Google Scholar]
  16. 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]
  17. Linke M. J., Smulian A. G., Stringer J. R., Walzer P. D. Characterization of multiple unique cDNAs encoding the major surface glycoprotein of rat-derived Pneumocystis carinii. Parasitol Res. 1994;80(6):478–486. doi: 10.1007/BF00932694. [DOI] [PubMed] [Google Scholar]
  18. Linke M. J., Smulian A. G., Yoshihara P., Walzer P. D. Production and characterization of monoclonal antibodies specific for the major surface glycoprotein of Pneumocystis carinii. J Eukaryot Microbiol. 1994 Sep-Oct;41(5):99S–100S. [PubMed] [Google Scholar]
  19. Liu Y., Leibowitz M. J. Variation and in vitro splicing of group I introns in rRNA genes of Pneumocystis carinii. Nucleic Acids Res. 1993 May 25;21(10):2415–2421. doi: 10.1093/nar/21.10.2415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lu J. J., Bartlett M. S., Shaw M. M., Queener S. F., Smith J. W., Ortiz-Rivera M., Leibowitz M. J., Lee C. H. Typing of Pneumocystis carinii strains that infect humans based on nucleotide sequence variations of internal transcribed spacers of rRNA genes. J Clin Microbiol. 1994 Dec;32(12):2904–2912. doi: 10.1128/jcm.32.12.2904-2912.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pixley F. J., Wakefield A. E., Banerji S., Hopkin J. M. Mitochondrial gene sequences show fungal homology for Pneumocystis carinii. Mol Microbiol. 1991 Jun;5(6):1347–1351. doi: 10.1111/j.1365-2958.1991.tb00781.x. [DOI] [PubMed] [Google Scholar]
  22. Pottratz S. T., Paulsrud J., Smith J. S., Martin W. J., 2nd Pneumocystis carinii attachment to cultured lung cells by pneumocystis gp 120, a fibronectin binding protein. J Clin Invest. 1991 Aug;88(2):403–407. doi: 10.1172/JCI115318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Smulian A. G., Stringer J. R., Linke M. J., Walzer P. D. Isolation and characterization of a recombinant antigen of Pneumocystis carinii. Infect Immun. 1992 Mar;60(3):907–915. doi: 10.1128/iai.60.3.907-915.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Smulian A. G., Sullivan D. W., Linke M. J., Halsey N. A., Quinn T. C., MacPhail A. P., Hernandez-Avila M. A., Hong S. T., Walzer P. D. Geographic variation in the humoral response to Pneumocystis carinii. J Infect Dis. 1993 May;167(5):1243–1247. doi: 10.1093/infdis/167.5.1243. [DOI] [PubMed] [Google Scholar]
  25. Stringer J. R., Stringer S. L., Zhang J., Baughman R., Smulian A. G., Cushion M. T. Molecular genetic distinction of Pneumocystis carinii from rats and humans. J Eukaryot Microbiol. 1993 Nov-Dec;40(6):733–741. doi: 10.1111/j.1550-7408.1993.tb04468.x. [DOI] [PubMed] [Google Scholar]
  26. Stringer S. L., Hong S. T., Giuntoli D., Stringer J. R. Repeated DNA in Pneumocystis carinii. J Clin Microbiol. 1991 Jun;29(6):1194–1201. doi: 10.1128/jcm.29.6.1194-1201.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stringer S. L., Stringer J. R., Blase M. A., Walzer P. D., Cushion M. T. Pneumocystis carinii: sequence from ribosomal RNA implies a close relationship with fungi. Exp Parasitol. 1989 May;68(4):450–461. doi: 10.1016/0014-4894(89)90130-6. [DOI] [PubMed] [Google Scholar]
  28. Sunkin S. M., Stringer S. L., Stringer J. R. A tandem repeat of rat-derived Pneumocystis carinii genes encoding the major surface glycoprotein. J Eukaryot Microbiol. 1994 May-Jun;41(3):292–300. doi: 10.1111/j.1550-7408.1994.tb01509.x. [DOI] [PubMed] [Google Scholar]
  29. 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. doi: 10.1128/iai.57.5.1363-1368.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Vasquez J., Smulian A. G., Linke M. J., Cushion M. T. Antigenic differences among genetically distinct types of rat-derived Pneumocystis carinii. J Eukaryot Microbiol. 1994 Sep-Oct;41(5):114S–114S. [PubMed] [Google Scholar]
  32. Wada M., Sunkin S. M., Stringer J. R., Nakamura Y. Antigenic variation by positional control of major surface glycoprotein gene expression in Pneumocystis carinii. J Infect Dis. 1995 Jun;171(6):1563–1568. doi: 10.1093/infdis/171.6.1563. [DOI] [PubMed] [Google Scholar]
  33. Wakefield A. E., Peters S. E., Banerji S., Bridge P. D., Hall G. S., Hawksworth D. L., Guiver L. A., Allen A. G., Hopkin J. M. Pneumocystis carinii shows DNA homology with the ustomycetous red yeast fungi. Mol Microbiol. 1992 Jul;6(14):1903–1911. doi: 10.1111/j.1365-2958.1992.tb01363.x. [DOI] [PubMed] [Google Scholar]
  34. Walzer P. D., Kim C. K., Linke M. J., Pogue C. L., Huerkamp M. J., Chrisp C. E., Lerro A. V., Wixson S. K., Hall E., Shultz L. D. Outbreaks of Pneumocystis carinii pneumonia in colonies of immunodeficient mice. Infect Immun. 1989 Jan;57(1):62–70. doi: 10.1128/iai.57.1.62-70.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Weinberg G. A., Dykstra C. C., Durant P. J., Cushion M. T. Chromosomal localization of 20 genes to five distinct pulsed field gel karyotypic forms of rat Pneumocystis carinii. J Eukaryot Microbiol. 1994 Sep-Oct;41(5):117S–117S. [PubMed] [Google Scholar]
  37. Ypma-Wong M. F., Fonzi W. A., Sypherd P. S. Fungus-specific translation elongation factor 3 gene present in Pneumocystis carinii. Infect Immun. 1992 Oct;60(10):4140–4145. doi: 10.1128/iai.60.10.4140-4145.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES