Skip to main content
PMC home page
Clinical and Diagnostic Laboratory Immunology logoLink to Clinical and Diagnostic Laboratory Immunology
. 1997 May;4(3):279–284. doi: 10.1128/cdli.4.3.279-284.1997

Antibody and delayed-type hypersensitivity responses to Ochrobactrum anthropi cytosolic and outer membrane antigens in infections by smooth and rough Brucella spp.

J Velasco 1, R Díaz 1, M J Grilló 1, M Barberán 1, C Marín 1, J M Blasco 1, I Moriyón 1
PMCID: PMC170519  PMID: 9144364

Abstract

Immunological cross-reactions between Brucella spp. and Ochrobactrum anthropi were investigated in animals and humans naturally infected by Brucella spp. and in experimentally infected rams (Brucella ovis infected), rabbits (Brucella melitensis infected), and mice (B. melitensis and Brucella abortus infected). In the animals tested, O. anthropi cytosolic proteins evoked a delayed-type hypersensitivity reaction of a frequency and intensity similar to that observed with B. melitensis brucellin. O. anthropi cytosolic proteins also reacted in gel precipitation tests with antibodies in sera from Brucella natural hosts with a frequency similar to that observed with B. melitensis proteins, and absorption experiments and immunoblotting showed antibodies to both Brucella-specific proteins and proteins common to Brucella and O. anthropi. No antibodies to O. anthropi cytosolic proteins were detected in the sera of Brucella-free hosts. Immunoblotting with sera of Brucella-infected sheep and goats showed immunoglobulin G (IgG) to Brucella group 3 outer membrane proteins and to O. anthropi proteins of similar molecular weight. No IgG to the O-specific polysaccharide of O. anthropi lipopolysaccharide was detected in the sera of Brucella-infected hosts. The sera of sheep, goats, and rabbits infected with B. melitensis contained IgG to O. anthropi rough lipopolysaccharide and lipid A, and B. ovis and O. anthropi rough lipopolysaccharides showed equal reactivities with IgG in the sera of B. ovis-infected rams. The findings show that the immunoresponse of Brucella-infected hosts to protein antigens is not necessarily specific for brucellae and suggest that the presence of O. anthropi or some related bacteria explains the previously described reactivities to Brucella rough lipopolysaccharide and outer membrane proteins in healthy animals.

Full Text

The Full Text of this article is available as a PDF (873.3 KB).

Selected References

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

  1. Alonso-Urmeneta B., Moriyón I., Díaz R., Blasco J. M. Enzyme-linked immunosorbent assay with Brucella native hapten polysaccharide and smooth lipopolysaccharide. J Clin Microbiol. 1988 Dec;26(12):2642–2646. doi: 10.1128/jcm.26.12.2642-2646.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baldi P. C., Giambartolomei G. H., Goldbaum F. A., Abdón L. F., Velikovsky C. A., Kittelberger R., Fossati C. A. Humoral immune response against lipopolysaccharide and cytoplasmic proteins of Brucella abortus in cattle vaccinated with B. abortus S19 or experimentally infected with Yersinia enterocolitica serotype 0:9. Clin Diagn Lab Immunol. 1996 Jul;3(4):472–476. doi: 10.1128/cdli.3.4.472-476.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bizet C., Bizet J. Sensibilité comparée de Ochrobactrum anthropi, Agrobacterium tumefaciens, Alcaligenes faecalis, Alcaligenes denitrificans subsp. denitrificans, Alcaligenes denitrificans subsp. xylosidans et Bordetella bronchiseptica vis-à-vis de 35 antibiotiques dont 17 beta-lactamines. Pathol Biol (Paris) 1995 Apr;43(4):258–263. [PubMed] [Google Scholar]
  4. Blasco J. M., Marín C., Jiménez de Bagués M., Barberán M., Hernández A., Molina L., Velasco J., Díaz R., Moriyón I. Evaluation of allergic and serological tests for diagnosing Brucella melitensis infection in sheep. J Clin Microbiol. 1994 Aug;32(8):1835–1840. doi: 10.1128/jcm.32.8.1835-1840.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cieslak T. J., Robb M. L., Drabick C. J., Fischer G. W. Catheter-associated sepsis caused by Ochrobactrum anthropi: report of a case and review of related nonfermentative bacteria. Clin Infect Dis. 1992 Apr;14(4):902–907. doi: 10.1093/clinids/14.4.902. [DOI] [PubMed] [Google Scholar]
  6. Cloeckaert A., Kerkhofs P., Limet J. N. Antibody response to Brucella outer membrane proteins in bovine brucellosis: immunoblot analysis and competitive enzyme-linked immunosorbent assay using monoclonal antibodies. J Clin Microbiol. 1992 Dec;30(12):3168–3174. doi: 10.1128/jcm.30.12.3168-3174.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Diaz R., Jones L. M., Leong D., Wilson J. B. Surface antigens of smooth brucellae. J Bacteriol. 1968 Oct;96(4):893–901. doi: 10.1128/jb.96.4.893-901.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Diaz R., Jones L. M., Wilson J. B. Antigenic relationship of the gram-negative organism causing canine abortion to smooth and rough brucellae. J Bacteriol. 1968 Feb;95(2):618–624. doi: 10.1128/jb.95.2.618-624.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dubray G. Progrès récents sur la biochimie et les propriétés biologiques des antigènes de Brucella. Dev Biol Stand. 1984;56:131–150. [PubMed] [Google Scholar]
  10. Dáz R., Maravi-Poma E., Rivero A. Comparison of counter-immunoelectrophoresis with other serological tests in the diagnosis of human brucellosis. Bull World Health Organ. 1976;53(4):417–424. [PMC free article] [PubMed] [Google Scholar]
  11. Díaz-Aparicio E., Aragón V., Marín C., Alonso B., Font M., Moreno E., Pérez-Ortiz S., Blasco J. M., Díaz R., Moriyón I. Comparative analysis of Brucella serotype A and M and Yersinia enterocolitica O:9 polysaccharides for serological diagnosis of brucellosis in cattle, sheep, and goats. J Clin Microbiol. 1993 Dec;31(12):3136–3141. doi: 10.1128/jcm.31.12.3136-3141.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Díaz-Aparicio E., Marín C., Alonso-Urmeneta B., Aragón V., Pérez-Ortiz S., Pardo M., Blasco J. M., Díaz R., Moriyón I. Evaluation of serological tests for diagnosis of Brucella melitensis infection of goats. J Clin Microbiol. 1994 May;32(5):1159–1165. doi: 10.1128/jcm.32.5.1159-1165.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Díaz R., Bosseray N. Estudio de las relaciones antigénicas entre Yersinia enterocolitica serotipo 9 y otras especies bacterianas gram-negavtivas. Microbiol Esp. 1974 Jan-Mar;27(1):1–14. [PubMed] [Google Scholar]
  14. Ezzedine H., Mourad M., Van Ossel C., Logghe C., Squifflet J. P., Renault F., Wauters G., Gigi J., Wilmotte L., Haxhe J. J. An outbreak of Ochrobactrum anthropi bacteraemia in five organ transplant patients. J Hosp Infect. 1994 May;27(1):35–42. doi: 10.1016/0195-6701(94)90066-3. [DOI] [PubMed] [Google Scholar]
  15. Ficapal A., Alonso-Urmeneta B., Velasco J., Moriyón I., Blasco J. M. Diagnosis of Brucella ovis infection of rams with an ELISA using protein G as conjugate. Vet Rec. 1995 Aug 5;137(6):145–147. doi: 10.1136/vr.137.6.145. [DOI] [PubMed] [Google Scholar]
  16. Galanos C., Lüderitz O., Westphal O. A new method for the extraction of R lipopolysaccharides. Eur J Biochem. 1969 Jun;9(2):245–249. doi: 10.1111/j.1432-1033.1969.tb00601.x. [DOI] [PubMed] [Google Scholar]
  17. Gamazo C., Winter A. J., Moriyón I., Riezu-Boj J. I., Blasco J. M., Díaz R. Comparative analyses of proteins extracted by hot saline or released spontaneously into outer membrane blebs from field strains of Brucella ovis and Brucella melitensis. Infect Immun. 1989 May;57(5):1419–1426. doi: 10.1128/iai.57.5.1419-1426.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Goldbaum F. A., Rubbi C. P., Wallach J. C., Miguel S. E., Baldi P. C., Fossati C. A. Differentiation between active and inactive human brucellosis by measuring antiprotein humoral immune responses. J Clin Microbiol. 1992 Mar;30(3):604–607. doi: 10.1128/jcm.30.3.604-607.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gransden W. R., Eykyn S. J. Seven cases of bacteremia due to Ochrobactrum anthropi. Clin Infect Dis. 1992 Dec;15(6):1068–1069. doi: 10.1093/clind/15.6.1068. [DOI] [PubMed] [Google Scholar]
  20. Hemmen F., Weynants V., Scarcez T., Letesson J. J., Saman E. Cloning and sequence analysis of a newly identified Brucella abortus gene and serological evaluation of the 17-kilodalton antigen that it encodes. Clin Diagn Lab Immunol. 1995 May;2(3):263–267. doi: 10.1128/cdli.2.3.263-267.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Iannelli D., Diaz R., Bettini T. M. Identification of Brucella abortus antibodies in cattle serum by single radial diffusion. J Clin Microbiol. 1976 Feb;3(2):203–205. doi: 10.1128/jcm.3.2.203-205.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kern W. V., Oethinger M., Kaufhold A., Rozdzinski E., Marre R. Ochrobactrum anthropi bacteremia: report of four cases and short review. Infection. 1993 Sep-Oct;21(5):306–310. doi: 10.1007/BF01712451. [DOI] [PubMed] [Google Scholar]
  23. Leong D., Diaz R., Milner K., Rudbach J., Wilson J. B. Some structural and biological properties of Brucella endotoxin. Infect Immun. 1970 Feb;1(2):174–182. doi: 10.1128/iai.1.2.174-182.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lin J., Adams L. G., Ficht T. A. Characterization of the heat shock response in Brucella abortus and isolation of the genes encoding the GroE heat shock proteins. Infect Immun. 1992 Jun;60(6):2425–2431. doi: 10.1128/iai.60.6.2425-2431.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Moreno E., Jones L. M., Berman D. T. Immunochemical characterization of rough Brucella lipopolysaccharides. Infect Immun. 1984 Mar;43(3):779–782. doi: 10.1128/iai.43.3.779-782.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Moriyon I., Berman D. T. Effects of nonionic, ionic, and dipolar ionic detergents and EDTA on the Brucella cell envelope. J Bacteriol. 1982 Nov;152(2):822–828. doi: 10.1128/jb.152.2.822-828.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Riezu-Boj J. I., Moriyón I., Blasco J. M., Gamazo C., Díaz R. Antibody response to Brucella ovis outer membrane proteins in ovine brucellosis. Infect Immun. 1990 Feb;58(2):489–494. doi: 10.1128/iai.58.2.489-494.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rojas N., Freer E., Weintraub A., Ramirez M., Lind S., Moreno E. Immunochemical identification of Brucella abortus lipopolysaccharide epitopes. Clin Diagn Lab Immunol. 1994 Mar;1(2):206–213. doi: 10.1128/cdli.1.2.206-213.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Romero C., Gamazo C., Pardo M., López-Goñi I. Specific detection of Brucella DNA by PCR. J Clin Microbiol. 1995 Mar;33(3):615–617. doi: 10.1128/jcm.33.3.615-617.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Roop R. M., 2nd, Fletcher T. W., Sriranganathan N. M., Boyle S. M., Schurig G. G. Identification of an immunoreactive Brucella abortus HtrA stress response protein homolog. Infect Immun. 1994 Mar;62(3):1000–1007. doi: 10.1128/iai.62.3.1000-1007.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rossetti O. L., Arese A. I., Boschiroli M. L., Cravero S. L. Cloning of Brucella abortus gene and characterization of expressed 26-kilodalton periplasmic protein: potential use for diagnosis. J Clin Microbiol. 1996 Jan;34(1):165–169. doi: 10.1128/jcm.34.1.165-169.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Salih-Alj Debbarh H., Cloeckaert A., Bézard G., Dubray G., Zygmunt M. S. Enzyme-linked immunosorbent assay with partially purified cytosoluble 28-kilodalton protein for serological differentiation between Brucella melitensis-infected and B. melitensis Rev.1-vaccinated sheep. Clin Diagn Lab Immunol. 1996 May;3(3):305–308. doi: 10.1128/cdli.3.3.305-308.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schurig G. G., Roop R. M., 2nd, Bagchi T., Boyle S., Buhrman D., Sriranganathan N. Biological properties of RB51; a stable rough strain of Brucella abortus. Vet Microbiol. 1991 Jul;28(2):171–188. doi: 10.1016/0378-1135(91)90091-s. [DOI] [PubMed] [Google Scholar]
  34. Tabatabai L. B., Hennager S. G. Cattle serologically positive for Brucella abortus have antibodies to B. abortus Cu-Zn superoxide dismutase. Clin Diagn Lab Immunol. 1994 Sep;1(5):506–510. doi: 10.1128/cdli.1.5.506-510.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tall B. D., Williams H. N., George K. S., Gray R. T., Walch M. Bacterial succession within a biofilm in water supply lines of dental air-water syringes. Can J Microbiol. 1995 Jul;41(7):647–654. doi: 10.1139/m95-088. [DOI] [PubMed] [Google Scholar]
  36. Velasco J., Moll H., Vinogradov E. V., Moriyón I., Zähringer U. Determination of the O-specific polysaccharide structure in the lipopolysaccharide of Ochrobactrum anthropi LMG 3331. Carbohydr Res. 1996 Jun 7;287(1):123–126. doi: 10.1016/0008-6215(96)00068-7. [DOI] [PubMed] [Google Scholar]
  37. Zygmunt M. S., Cloeckaert A., Dubray G. Brucella melitensis cell envelope protein and lipopolysaccharide epitopes involved in humoral immune responses of naturally and experimentally infected sheep. J Clin Microbiol. 1994 Oct;32(10):2514–2522. doi: 10.1128/jcm.32.10.2514-2522.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Zygmunt M. S., Gilbert F. B., Dubray G. Purification, characterization, and seroactivity of a 20-kilodalton Brucella protein antigen. J Clin Microbiol. 1992 Oct;30(10):2662–2667. doi: 10.1128/jcm.30.10.2662-2667.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Clinical and Diagnostic Laboratory Immunology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES