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. 1992 May;30(5):1183–1188. doi: 10.1128/jcm.30.5.1183-1188.1992

Isolation and purification of two immunodominant antigens from Nocardia brasiliensis.

L Vera-Cabrera 1, M C Salinas-Carmona 1, O Welsh 1, M A Rodriguez 1
PMCID: PMC265247  PMID: 1583118

Abstract

Two immunogenic proteins from a crude extract of Nocardia brasiliensis were purified to homogeneity. A 61-kDa protein (P61) was isolated from a 50% ammonium sulfate precipitate in two steps. Initially, P61 was obtained by electroelution in a 10% nondenatured preparative polyacrylamide gel electrophoresis (PAGE). In a second step, the eluate from the nondenatured gel was run in a 12% sodium dodecyl sulfate (SDS) preparative polyacrylamide gel. After elution, a single band was demonstrated by SDS-PAGE and Western blot (immunoblot). Also, a 24-kDa immunogenic protein (P24) was isolated by gel filtration in a Sephadex G-100 column and then by electroelution in a 12% nondenatured polyacrylamide gel. In a previous paper, we showed by Western blot assays that these proteins are recognized by the sera of mycetoma patients and not by sera from mycobacterial-infected or healthy individuals. We consider these proteins to be good candidates for the study of the host-parasite relationship in nocardial infections. The possible clinical application of these purified antigens in a serological diagnosis is discussed.

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Selected References

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  1. Angeles A. M., Sugar A. M. Identification of a common immunodominant protein in culture filtrates of three Nocardia species and use in etiologic diagnosis of mycetoma. J Clin Microbiol. 1987 Dec;25(12):2278–2280. doi: 10.1128/jcm.25.12.2278-2280.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Angeles A. M., Sugar A. M. Rapid diagnosis of nocardiosis with an enzyme immunoassay. J Infect Dis. 1987 Feb;155(2):292–296. doi: 10.1093/infdis/155.2.292. [DOI] [PubMed] [Google Scholar]
  3. Beaman B. L., Burnside J., Edwards B., Causey W. Nocardial infections in the United States, 1972-1974. J Infect Dis. 1976 Sep;134(3):286–289. doi: 10.1093/infdis/134.3.286. [DOI] [PubMed] [Google Scholar]
  4. Beaman B. L., Maslan S. Effect of cyclophosphamide on experimental Nocardia asteroides infection in mice. Infect Immun. 1977 Jun;16(3):995–1004. doi: 10.1128/iai.16.3.995-1004.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blumer S. O., Kaufman L. Microimmunodiffusion test for nocardiosis. J Clin Microbiol. 1979 Sep;10(3):308–312. doi: 10.1128/jcm.10.3.308-312.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Boiron P., Provost F. Use of partially purified 54-kilodalton antigen for diagnosis of nocardiosis by Western blot (immunoblot) assay. J Clin Microbiol. 1990 Feb;28(2):328–331. doi: 10.1128/jcm.28.2.328-331.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  8. Buot G., Lavalle P., Mariat F., Suchil P. Etude épidémiologique des mycétomes au Mexique. A propos de 502 cas. Bull Soc Pathol Exot Filiales. 1987;80(3):329–339. [PubMed] [Google Scholar]
  9. Deem R. L., Beaman B. L., Gershwin M. E. Adoptive transfer of immunity to Nocardia asteroides in nude mice. Infect Immun. 1982 Dec;38(3):914–920. doi: 10.1128/iai.38.3.914-920.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Filice G. A., Beaman B. L., Remington J. S. Effects of activated macrophages on Nacardia asteroides. Infect Immun. 1980 Feb;27(2):643–649. doi: 10.1128/iai.27.2.643-649.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jiménez T., Díaz A. M., Zlotnik H. Monoclonal antibodies to Nocardia asteroides and Nocardia brasiliensis antigens. J Clin Microbiol. 1990 Jan;28(1):87–91. doi: 10.1128/jcm.28.1.87-91.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kwapinski J. B., Kwapinski E. H., Dowler J., Horsman G. The phyloantigenic position of nocardiae revealed by examination of cytoplasmic antigens. Can J Microbiol. 1973 Aug;19(8):955–964. doi: 10.1139/m73-153. [DOI] [PubMed] [Google Scholar]
  13. 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]
  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. Ortiz-Ortiz L., Contreras M. F., Bujalil L. F. Cytoplasmic antigens from Nocardia elicting a specific delayed hypersensitivity. Infect Immun. 1972 Jun;5(6):879–882. doi: 10.1128/iai.5.6.879-882.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rico G., Ochoa R., Oliva A., Gonzalez-Mendoza A., Walker S. M., Ortiz-Ortiz L. Enhanced resistance to Nocardia brasiliensis infection in mice depleted of antigen-specific B cells. J Immunol. 1982 Oct;129(4):1688–1693. [PubMed] [Google Scholar]
  17. Ridell M. Immunodiffusion studies of some Nocardia strains. J Gen Microbiol. 1981 Mar;123(1):69–74. doi: 10.1099/00221287-123-1-69. [DOI] [PubMed] [Google Scholar]
  18. Stevens D. A., Pier A. C., Beaman B. L., Morozumi P. A., Lovett I. S., Houang E. T. Laboratory evaluation of an outbreak of nocardiosis in immunocompromised hosts. Am J Med. 1981 Dec;71(6):928–934. doi: 10.1016/0002-9343(81)90302-8. [DOI] [PubMed] [Google Scholar]
  19. Sugar A. M., Schoolnik G. K., Stevens D. A. Antibody response in human nocardiosis: identification of two immunodominant culture-filtrate antigens derived from Nocardia asteroides. J Infect Dis. 1985 May;151(5):895–901. doi: 10.1093/infdis/151.5.895. [DOI] [PubMed] [Google Scholar]
  20. Welsh O. Mycetoma. Current concepts in treatment. Int J Dermatol. 1991 Jun;30(6):387–398. doi: 10.1111/j.1365-4362.1991.tb03892.x. [DOI] [PubMed] [Google Scholar]
  21. el-Zaatari F. A., Reiss E., Yakrus M. A., Bragg S. L., Kaufman L. Monoclonal antibodies against isoelectrically focused Nocardia asteroides proteins characterized by the enzyme-linked immunoelectro-transfer blot method. Diagn Immunol. 1986;4(2):97–106. [PubMed] [Google Scholar]

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