Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1994 Dec;62(12):5361–5366. doi: 10.1128/iai.62.12.5361-5366.1994

Brucella ribosomal protein L7/L12 is a major component in the antigenicity of brucellin INRA for delayed-type hypersensitivity in brucella-sensitized guinea pigs.

G Bachrach 1, M Banai 1, S Bardenstein 1, G Hoida 1, A Genizi 1, H Bercovier 1
PMCID: PMC303276  PMID: 7960115

Abstract

A delayed-type hypersensitivity (DTH) reaction in the course of brucellosis in humans and animals can be revealed by the brucellin INRA (Brucellergen) skin test. Brucellergen is composed of more than 20 proteins of different molecular weights. A 12-kDa protein eliciting DTH in Brucella melitensis Rev1-sensitized guinea pigs was found to be a significant component for the allergenic properties of Brucellergen. Sequencing of the gene encoding this protein identified it as the L7/L12 ribosomal protein. The L7/L12 gene of B. melitensis was amplified by PCR and subcloned in the Escherichia coli pQE30 plasmid. The resulting recombinant protein did not produce a DTH reaction in sensitized animals. It was used to raise specific antibodies in a rabbit. Affinity chromatography with these antibodies was used to isolate a single protein from Brucellergen and from B. melitensis cytosol preparations which produced a DTH reaction in guinea pigs sensitized with B. melitensis Rev1. N-terminal amino acid sequencing of the protein confirmed that it was the L7/L12 ribosomal protein. This is the first complete report on the involvement of a defined bacterial ribosomal protein in the DTH response of animals infected with intracellularly multiplying bacteria.

Full text

PDF
5361

Images in this article

Selected References

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

  1. Bachrach G., Bar-Nir D., Banai M., Bercovier H. Identification and nucleotide sequence of Brucella melitensis L7/L12 ribosomal protein. FEMS Microbiol Lett. 1994 Jul 15;120(3):237–240. doi: 10.1111/j.1574-6968.1994.tb07039.x. [DOI] [PubMed] [Google Scholar]
  2. Baker R. E., Hill W. E., Larson C. L. Delayed hypersensitivity reactions provoked by ribosomes from acid-fast bacilli. I. Ribosomal isolation, characterization, delayed hypersensitivity, and specificity. Infect Immun. 1972 Sep;6(3):258–265. doi: 10.1128/iai.6.3.258-265.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baker R. E., Hill W. E., Larson C. L. Ribosomes of acid-fast bacilli: immunogenicity, serology, and in vitro correlates of delayed hypersensitivity. Infect Immun. 1973 Aug;8(2):236–244. doi: 10.1128/iai.8.2.236-244.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bhongbhibhat N., Elberg S., Chen T. H. Characterization of brucella skin-test antigens. J Infect Dis. 1970 Jul-Aug;122(1):70–82. doi: 10.1093/infdis/122.1-2.70. [DOI] [PubMed] [Google Scholar]
  5. 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: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  6. Brooks-Worrell B. M., Splitter G. A. Antigens of Brucella abortus S19 immunodominant for bovine lymphocytes as identified by one- and two-dimensional cellular immunoblotting. Infect Immun. 1992 Jun;60(6):2459–2464. doi: 10.1128/iai.60.6.2459-2464.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brooks-Worrell B. M., Splitter G. A. Sodium dodecyl sulfate- and salt-extracted antigens from various Brucella species induce proliferation of bovine lymphocytes. Infect Immun. 1992 May;60(5):2136–2138. doi: 10.1128/iai.60.5.2136-2138.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Corbel M. J. The immunogenic activity of ribosomal fractions derived from Brucella abortus. J Hyg (Lond) 1976 Feb;76(1):65–74. doi: 10.1017/s0022172400054954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dohoo I. R., Wright P. F., Ruckerbauer G. M., Samagh B. S., Robertson F. J., Forbes L. B. A comparison of five serological tests for bovine brucellosis. Can J Vet Res. 1986 Oct;50(4):485–493. [PMC free article] [PubMed] [Google Scholar]
  10. Fensterbank R. Diagnostic allergique de la brucellose bovine. 2. Utilisation du test allergique dans les troupeaux infectés. Ann Rech Vet. 1977;8(2):195–201. [PubMed] [Google Scholar]
  11. Fensterbank R., Pardon P. Diagnostic allergique de la brucellose bovine. 1. Conditions d'utilisation d'un allergène protéique purifié: la brucelline. Ann Rech Vet. 1977;8(2):187–193. [PubMed] [Google Scholar]
  12. Gregory R. L. Microbial ribosomal vaccines. Rev Infect Dis. 1986 Mar-Apr;8(2):208–217. doi: 10.1093/clinids/8.2.208. [DOI] [PubMed] [Google Scholar]
  13. Jones L. M., Berman D. T. Antibody-mediated and delayed-type hypersensitivity reactions to Brucella skin test antigens in guinea pigs. Infect Immun. 1975 Feb;11(2):360–364. doi: 10.1128/iai.11.2.360-364.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jones L. M., Diaz R., Taylor A. G. Characterization of allergens prepared from smooth and rough strains of Brucella melitensis. Br J Exp Pathol. 1973 Oct;54(5):492–508. [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Loge R. V., Hill W. E., Baker R. E., Larson C. L. Delayed hypersensitivity reactions provoked by ribosomes from acid-fast bacilli: physical characteristics and immunological aspects of core ribosomal proteins from Mycobacterium smegmatis. Infect Immun. 1974 Mar;9(3):489–496. doi: 10.1128/iai.9.3.489-496.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Oliveira S. C., Zhu Y., Splitter G. Sequences of the rplJL operon containing the L10 and L7/L12 genes from Brucella abortus. Gene. 1994 Mar 11;140(1):137–138. doi: 10.1016/0378-1119(94)90744-7. [DOI] [PubMed] [Google Scholar]
  18. Ortiz-Ortiz L., Solarolo E. B., Bojalil L. F. Delayed hypersensitivity to ribosomal protein from BCG. J Immunol. 1971 Oct;107(4):1022–1026. [PubMed] [Google Scholar]
  19. Stemshorn B. W. Recent progress in the diagnosis of brucellosis. Dev Biol Stand. 1984;56:325–340. [PubMed] [Google Scholar]
  20. Sterne M., Trim G., Broughton E. S. Immunisation of laboratory animals and cattle with non-agglutinogenic extracts of Brucella abortus strain 45-20. J Med Microbiol. 1971 May;4(2):185–194. doi: 10.1099/00222615-4-2-185. [DOI] [PubMed] [Google Scholar]
  21. Tantimavanich S., Nagai S., Nomaguchi H., Kinomoto M., Ohara N., Yamada T. Immunological properties of ribosomal proteins from Mycobacterium bovis BCG. Infect Immun. 1993 Sep;61(9):4005–4007. doi: 10.1128/iai.61.9.4005-4007.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Terhorst C., Wittmann-Liebold B., Möller W. 50-S ribosomal proteins. Peptide studies on two acidic proteins, A 1 and A 2 , isolated from 50-S ribosomes of Escherichia coli. Eur J Biochem. 1972 Jan 31;25(1):13–19. doi: 10.1111/j.1432-1033.1972.tb01661.x. [DOI] [PubMed] [Google Scholar]
  23. Thomas D. W., Weiss E. Response of mice to injection of ribosomal fraction from group B Neisseria meningitidis. Infect Immun. 1972 Sep;6(3):355–363. doi: 10.1128/iai.6.3.355-363.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Thompson H. C., Snyder I. S. Protection against pneumococcal infection by a ribosomal preparation. Infect Immun. 1971 Jan;3(1):16–23. doi: 10.1128/iai.3.1.16-23.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Venneman M. R., Bigley N. J. Isolation and partial characterization of an immunogenic moiety obtained from Salmonella typhimurium. J Bacteriol. 1969 Oct;100(1):140–148. doi: 10.1128/jb.100.1.140-148.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. YOUMANS A. S., YOUMANS G. P. IMMUNOGENIC ACTIVITY OF A RIBOSOMAL FRACTION OBTAINED FROM MYCOBACTERIUM TUBERCULOSIS. J Bacteriol. 1965 May;89:1291–1298. doi: 10.1128/jb.89.5.1291-1298.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES