Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1992 Jul;60(7):2667–2671. doi: 10.1128/iai.60.7.2667-2671.1992

Isolation and antigenicity of a 45-kilodalton Paracoccidioides brasiliensis immunodominant antigen.

M F Ferreira-da-Cruz 1, B Galvão-Castro 1, C Daniel-Ribeiro 1
PMCID: PMC257218  PMID: 1612736

Abstract

In the present study, we analyzed human antibody responses to Paracoccidioides brasiliensis cellular antigens by the immunoblot technique to identify specific cellular components and to investigate the existence of antigen profile differences among serological responses of paracoccidioidomycosis (PCM) patients. Among the 64 PCM serum samples analyzed, a relatively homogeneous immunoglobulin G response to P. brasiliensis antigens was observed. The polypeptide with a mass of 45 kDa was the most clinically important, since antibody to this antigen was detectable in 90.6% of PCM patients studied and the six individuals who did not produce antibody were either at the end of treatment or in the posttherapy period and had shown clinical recovery. These facts suggested that the presence of this antibody may be an indicator of active disease. The 45-kDa antigen was also the most specific antigen of the PCM humoral immune response, since it reacted with only 2 of 79 (2.5%) heterologous serum samples tested: 1 histoplasmosis case and 1 tuberculosis case. This polypeptide was isolated from gels by electroelution and, when tested by an immunoradiometric assay and immunoblotting, maintained its reactivity with PCM sera and also with anti-P. brasiliensis polyclonal antibodies raised in rabbits at the same sensitivity levels as those obtained in immunoblotting with a crude antigen. Since in our assays the 45-kDa polypeptide was the major P. brasiliensis antigen and seemed to be specific for PCM, its use in alternative diagnostic methods is promising, especially in patients suspected of having the juvenile clinical form of PCM often associated with negative double-immunodiffusion results.

Full text

PDF
2667

Images in this article

Selected References

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

  1. Blumer S. O., Jalbert M., Kaufman L. Rapid and reliable method for production of a specific Paracoccidioides brasiliensis immunodiffusion test antigen. J Clin Microbiol. 1984 Mar;19(3):404–407. doi: 10.1128/jcm.19.3.404-407.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bongertz V. Eluiço de proteínas de gel de poliacrilamida: descriço de metodologia simples e econômica. Rev Inst Med Trop Sao Paulo. 1989 Jan-Feb;31(1):44–47. doi: 10.1590/s0036-46651989000100009. [DOI] [PubMed] [Google Scholar]
  3. Burgos L. C., Cano L. E., Restrepo A. Purificación de antígenos somáticos del Paracoccidioides brasiliensis. Estudio preliminar. Rev Inst Med Trop Sao Paulo. 1985 Mar-Apr;27(2):76–81. doi: 10.1590/s0036-46651985000200003. [DOI] [PubMed] [Google Scholar]
  4. Camargo Z. P., Unterkircher C., Travassos L. R. Identification of antigenic polypeptides of Paracoccidioides brasiliensis by immunoblotting. J Med Vet Mycol. 1989;27(6):407–412. [PubMed] [Google Scholar]
  5. Casotto M. Characterization of the cellular antigens of Paracoccidioides brasiliensis yeast form. J Clin Microbiol. 1990 Jun;28(6):1188–1193. doi: 10.1128/jcm.28.6.1188-1193.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Casotto M., Paris S., Camargo Z. P. Antigens of diagnostic value in three isolates of Paracoccidioides brasiliensis. J Med Vet Mycol. 1991;29(4):243–253. doi: 10.1080/02681219180000361. [DOI] [PubMed] [Google Scholar]
  7. De Camargo Z., Unterkircher C., Campoy S. P., Travassos L. R. Production of Paracoccidioides brasiliensis exoantigens for immunodiffusion tests. J Clin Microbiol. 1988 Oct;26(10):2147–2151. doi: 10.1128/jcm.26.10.2147-2151.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ferreira-da-Cruz M. F., Francesconi-do-Vale A. C., Espinera M. C., Wanke B., Galvao-Castro B. Study of antibodies in paracoccidioidomycosis: follow-up of patients during and after treatment. J Med Vet Mycol. 1990;28(2):151–157. doi: 10.1080/02681219080000201. [DOI] [PubMed] [Google Scholar]
  9. Ferreira-da-Cruz M. F., Galvão-Castro B., Daniel-Ribeiro C. T. Sensitive immunoradiometric assay for the detection of Paracoccidioides brasiliensis antigens in human sera. J Clin Microbiol. 1991 Jun;29(6):1202–1205. doi: 10.1128/jcm.29.6.1202-1205.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ferreira-da-Cruz M. F., Galvão-Castro B., Wanke B. Producão e padronizaço dos antígenos de Paracoccidioides brasiliensis (Pb), Histoplasma capsulatum (Hc) e Aspergillus fumigatus (Af) para uso no imunodiagnóstico. Comparaço entre as técnicas de imunodifusão e imunoeletroosmoforese. Mem Inst Oswaldo Cruz. 1985 Jul-Sep;80(3):301–305. doi: 10.1590/s0074-02761985000300005. [DOI] [PubMed] [Google Scholar]
  11. Ferreire-da-Cruz M. F., Wanke B., Galvão-Castro B. Prevalence of paracoccidioidomycosis in hospitalized adults in Rio de Janeiro (RJ) Brazil. Mycopathologia. 1987 Jan;97(1):61–64. doi: 10.1007/BF00437332. [DOI] [PubMed] [Google Scholar]
  12. Fraker P. J., Speck J. C., Jr Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. Biochem Biophys Res Commun. 1978 Feb 28;80(4):849–857. doi: 10.1016/0006-291x(78)91322-0. [DOI] [PubMed] [Google Scholar]
  13. Kurup V. P., Greenberger P. A., Fink J. N. Antibody response to low-molecular-weight antigens of Aspergillus fumigatus in allergic bronchopulmonary aspergillosis. J Clin Microbiol. 1989 Jun;27(6):1312–1316. doi: 10.1128/jcm.27.6.1312-1316.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Matthews R. C., Burnie J. P., Tabaqchali S. Immunoblot analysis of the serological response in systemic candidosis. Lancet. 1984 Dec 22;2(8417-8418):1415–1418. doi: 10.1016/s0140-6736(84)91618-0. [DOI] [PubMed] [Google Scholar]
  16. Matthews R., Burnie J. P., Fox A., Tabaqchali S. Immunoblot analysis of serological responses in invasive aspergillosis. J Clin Pathol. 1985 Nov;38(11):1300–1303. doi: 10.1136/jcp.38.11.1300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McGowan K. L., Buckley H. R. Preparation and use of cytoplasmic antigens for the serodiagnosis of paracoccidioidomycosis. J Clin Microbiol. 1985 Jul;22(1):39–43. doi: 10.1128/jcm.22.1.39-43.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Puccia R., Schenkman S., Gorin P. A., Travassos L. R. Exocellular components of Paracoccidioides brasiliensis: identification of a specific antigen. Infect Immun. 1986 Jul;53(1):199–206. doi: 10.1128/iai.53.1.199-206.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Restrepo A., Moncada L. H. Characterization of the precipitin bands detected in the immunodiffusion test for paracoccidioidomycosis. Appl Microbiol. 1974 Jul;28(1):138–144. doi: 10.1128/am.28.1.138-144.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Restrepo M. A. La prueba de inmunodiffusion en el diagnostico de la paracoccidioidomicosis. Sabouraudia. 1966 Feb;4(4):223–230. [PubMed] [Google Scholar]
  21. Strockbine N. A., Largen M. T., Zweibel S. M., Buckley H. R. Identification and molecular weight characterization of antigens from Candida albicans that are recognized by human sera. Infect Immun. 1984 Feb;43(2):715–721. doi: 10.1128/iai.43.2.715-721.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Yarzabal L. A., Bout D., Naquira F., Fruit J., Andrieu S. Identification and purification of the specific antigen of Paracoccidioides brasiliensis responsible for immunoelectrophoretic band E. Sabouraudia. 1977 Mar;15(1):79–85. [PubMed] [Google Scholar]

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

RESOURCES