Skip to main content
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1994 Oct;38(10):2258–2265. doi: 10.1128/aac.38.10.2258

Use of beta-1,3-glucan-specific antibody to study the cyst wall of Pneumocystis carinii and effects of pneumocandin B0 analog L-733,560.

K H Nollstadt 1, M A Powles 1, H Fujioka 1, M Aikawa 1, D M Schmatz 1
PMCID: PMC284727  PMID: 7840554

Abstract

A new class of promising antipneumocystis agents, cyclic lipopeptide pneumocandin analogs, has been shown to effectively prevent Pneumocystis carinii cyst development in murine models. These compounds are believed to inhibit the biosynthesis of beta-1,3-glucan, a major constituent of the cell walls of various pathogenic fungi. However, all evidence of the presence of this polymer in P. carinii cysts is based on indirect methods. To address this, highly specific rabbit polyclonal antiserum was raised against a laminariheptaose-human transferrin hapten conjugate. This antiserum was used to demonstrate the presence of beta-1,3-glucan in alkaline extracts of P. carinii-infected rat lung tissue and to quantitate the degree of infection in this tissue as laminarin equivalents. The antiserum was also used to localize beta-1,3-glucan in P. carinii-infected rat lung tissue at the transmission electron microscopic level by immunogold labeling. High concentrations of beta-1,3-glucan were present in the electron-lucent layer of the P. carinii cyst wall, but beta-1,3-glucan was absent from intracystic bodies and trophozoites. Ultrastructural evaluation of lung tissue from P. carinii-infected rats treated with the pneumocandin analog L-733,560 demonstrated that the few cysts which remained are deformed, lack the translucent layer of the cyst wall, and contain minimal amounts of beta-1,3-glucan.

Full text

PDF
2258

Images in this article

Selected References

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

  1. Aikawa M., Atkinson C. T. Immunoelectron microscopy of parasites. Adv Parasitol. 1990;29:151–214. doi: 10.1016/s0065-308x(08)60106-2. [DOI] [PubMed] [Google Scholar]
  2. Bouffard F. A., Zambias R. A., Dropinski J. F., Balkovec J. M., Hammond M. L., Abruzzo G. K., Bartizal K. F., Marrinan J. A., Kurtz M. B., McFadden D. C. Synthesis and antifungal activity of novel cationic pneumocandin B(o) derivatives. J Med Chem. 1994 Jan 21;37(2):222–225. doi: 10.1021/jm00028a003. [DOI] [PubMed] [Google Scholar]
  3. Burke B. A., Good R. A. Pneumocystis carinii infection. 1973. Medicine (Baltimore) 1992 May;71(3):165–178. [PubMed] [Google Scholar]
  4. De Stefano J. A., Cushion M. T., Trinkle L. S., Walzer P. D. Lectins as probes to Pneumocystis carinii surface glycocomplexes. J Protozool. 1989 Jan-Feb;36(1):65S–66S. doi: 10.1111/j.1550-7408.1989.tb02704.x. [DOI] [PubMed] [Google Scholar]
  5. Dyer M., Volpe F., Delves C. J., Somia N., Burns S., Scaife J. G. Cloning and sequence of a beta-tubulin cDNA from Pneumocystis carinii: possible implications for drug therapy. Mol Microbiol. 1992 Apr;6(8):991–1001. doi: 10.1111/j.1365-2958.1992.tb02165.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. Edman U., Edman J. C., Lundgren B., Santi D. V. Isolation and expression of the Pneumocystis carinii thymidylate synthase gene. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6503–6507. doi: 10.1073/pnas.86.17.6503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gray G. R. The direct coupling of oligosaccharides to proteins and derivatized gels. Arch Biochem Biophys. 1974 Jul;163(1):426–428. doi: 10.1016/0003-9861(74)90495-0. [DOI] [PubMed] [Google Scholar]
  9. Hearn M. T., Bethell G. S., Ayers J. S., Hancock W. S. Application of 1,1'-carbonyldiimidazole-activated agarose for the purification of proteins. II. The use of an activated matrix devoid of additional charged groups for the purification of thyroid proteins. J Chromatogr. 1979 Dec 20;185:463–470. doi: 10.1016/s0021-9673(00)85622-8. [DOI] [PubMed] [Google Scholar]
  10. Kovacs J. A., Masur H. Pneumocystis carinii pneumonia: therapy and prophylaxis. J Infect Dis. 1988 Jul;158(1):254–259. doi: 10.1093/infdis/158.1.254. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Linke M. J., Walzer P. D. Analysis of a surface antigen of Pneumocystis carinii. J Protozool. 1989 Jan-Feb;36(1):60S–61S. doi: 10.1111/j.1550-7408.1989.tb02701.x. [DOI] [PubMed] [Google Scholar]
  13. Matsumoto Y., Matsuda S., Tegoshi T. Yeast glucan in the cyst wall of Pneumocystis carinii. J Protozool. 1989 Jan-Feb;36(1):21S–22S. doi: 10.1111/j.1550-7408.1989.tb02674.x. [DOI] [PubMed] [Google Scholar]
  14. Riis B., Rattan S. I., Clark B. F., Merrick W. C. Eukaryotic protein elongation factors. Trends Biochem Sci. 1990 Nov;15(11):420–424. doi: 10.1016/0968-0004(90)90279-k. [DOI] [PubMed] [Google Scholar]
  15. Schmatz D. M., Romancheck M. A., Pittarelli L. A., Schwartz R. E., Fromtling R. A., Nollstadt K. H., Vanmiddlesworth F. L., Wilson K. E., Turner M. J. Treatment of Pneumocystis carinii pneumonia with 1,3-beta-glucan synthesis inhibitors. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5950–5954. doi: 10.1073/pnas.87.15.5950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Williams D. J., Radding J. A., Dell A., Khoo K. H., Rogers M. E., Richards F. F., Armstrong M. Y. Glucan synthesis in Pneumocystis carinii. J Protozool. 1991 Jul-Aug;38(4):427–437. doi: 10.1111/j.1550-7408.1991.tb01382.x. [DOI] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES