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Infection and Immunity, November 2001, p. 7020-7028, Vol. 69, No. 11
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.11.7020-7028.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Minor Nucleotide Substitutions in the omp31 Gene of Brucella ovis Result in Antigenic Differences in the Major Outer Membrane Protein That It Encodes Compared to Those of the Other Brucella Species

Nieves Vizcaíno,1,* Reinhold Kittelberger,2 Axel Cloeckaert,3 Clara M. Marín,4 and Luis Fernández-Lago1

Departamento de Microbiología y Genética, Edificio Departmental, Universidad de Salamanca, 37007 Salamanca,1 and Unidad de Sanidad Animal, Servicio de Investigación Agroalimentaria, Diputación General de Aragón, 50080 Zaragoza,4 Spain; National Centre for Disease Investigation, Ministry of Agriculture and Forestry, Upper Hutt, New Zealand,2; and Station de Pathologie Aviaire et Parasitologie, Institut National de la Recherche Agronomique, Centre de Tours, 37380 Nouzilly, France3

Received 16 July 2001/Accepted 6 August 2001

The gene coding for the major outer membrane protein Omp31 was sequenced in five Brucella species and their biovars. Although the omp31 genes appeared to be highly conserved in the genus Brucella, nine nucleotide substitutions were detected in the gene of Brucella ovis compared to that of Brucella melitensis. As shown by differential binding properties of monoclonal antibodies (MAbs) to the two Brucella species, these nucleotide substitutions result in different antigenic properties of Omp31. The antigenic differences were also evidenced when sera from B. ovis-infected rams were tested by Western blotting with the recombinant B. melitensis or B. ovis Omp31 proteins. Twelve available sera reacted with recombinant B. ovis Omp31, but only four of them reacted with recombinant B. melitensis Omp31. These results validate previous evidence for the potential of Omp31 as a diagnostic antigen for B. ovis infection in rams and demonstrate that B. ovis Omp31, instead of B. melitensis Omp31, should be used to evaluate this point. The antigenic differences between the B. melitensis and B. ovis Omp31 proteins should also be taken into account when Omp31 is evaluated as a candidate for the development of subcellular vaccines against B. ovis infection. No reactivity against recombinant B. melitensis Omp31 was detected, by Western blotting, with sera from B. melitensis-infected sheep. Accordingly, Omp31 does not seem to be a good diagnostic antigen for B. melitensis infections in sheep. Two immunodominant regions were identified on the B. ovis Omp31 protein by using recombinant DNA techniques and specific MAbs. Sera from B. ovis-infected rams that reacted with the recombinant protein were tested by Western blotting against one of these immunodominant regions shown to be exposed at the bacterial surface. Only 4 of the 12 sera reacted, but with strong intensity.

* Corresponding author. Mailing address: Departamento de Microbiología y Genética, Edificio Departamental, Universidad de Salamanca, Avda. Campo Charro s/n, 37007 Salamanca, Spain. Phone: 34-923-294532. Fax: 34-923-224876. E-mail: vizcaino{at}www-micro.usal.es.

Infection and Immunity, November 2001, p. 7020-7028, Vol. 69, No. 11
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.11.7020-7028.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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