Characterization, occurrence, and molecular cloning of a 39-kilodalton Brucella abortus cytoplasmic protein immunodominant in cattle

• Published in: Infection and Immunity Vol. 65; no. 2; pp. 495 - 502
• Main Authors: Denoel, P.A. (Facultes Universitaires Notre-Dame de la Paix, Namur, Belgium.), Vo, T.K.O, Tibor, A, Weynants, V.E, Trunde, J.M, Dubray, G, Limet, J.N, Letesson, J.J
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.02.1997
• Subjects: Allergens - administration & dosage; Allergens - chemistry; Amino Acid Sequence; Animals; Antigens, Bacterial - administration & dosage; Antigens, Bacterial - genetics; Antigens, Bacterial - isolation & purification; Bacteriology; Base Sequence; Biological and medical sciences; BOVIN; BRUCELLA ABORTUS; Brucella abortus - chemistry; Brucella abortus - genetics; Brucella abortus - immunology; Cattle; Chromatography, Ion Exchange; CLONACION MOLECULAR; CLONAGE MOLECULAIRE; Cloning, Molecular; COMPOSICION QUIMICA; COMPOSITION CHIMIQUE; Cross Reactions; Cytoplasm - immunology; Female; Fundamental and applied biological sciences. Psychology; GANADO BOVINO; Genes, Bacterial; Hybridomas - chemistry; Hybridomas - immunology; Hypersensitivity, Delayed - etiology; Immunodominant Epitopes - administration & dosage; Immunodominant Epitopes - genetics; Immunodominant Epitopes - isolation & purification; Lymphocyte Activation; Microbiology; Molecular Sequence Data; Molecular Weight; Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains; SECUENCIA NUCLEOTIDICA; SEQUENCE NUCLEOTIDIQUE; Bovine; Nucleotide sequence; Host agent relation; Delayed hypersensitivity; Cellular immunity; Brucella abortus; Immunodominance; Infection; Proteins; Vertebrata; Mammalia; Brucellaceae; Animal; Bacteriosis; Bacteria; Brucellosis; Artiodactyla; Veterinary; Molecular cloning; Ungulata; Cytoplasm; Aminoacid sequence; Structural analysis
• ISSN: 0019-9567; 1098-5522
• DOI: 10.1128/iai.65.2.495-502.1997

Monoclonal antibodies and polyclonal antisera recognizing a 39-kDa protein (P39) of brucellin, a cytoplasmic extract from Brucella melitensis rough strain B115, were produced. The P39 was purified by anion-exchange chromatography. Eleven of fourteen Brucella-infected cows whose infections had been detected by the delayed-type hypersensitivity (DTH) test with brucellergen also developed a DTH reaction when purified P39 was used as the trigger. The T-cell proliferative responses to P39 of peripheral blood lymphocytes from Brucella-infected cows were also positive. None of the animals infected with other bacterial species that are presumed to induce immunological cross-reactions with Brucella spp. reacted to P39, either in DTH tests or in lymphocyte proliferation assays. A lambda gt11 genomic library of Brucella abortus was screened with a monoclonal antibody specific for P39, and the gene coding for this protein was subsequently isolated. The nucleotide sequence of the P39 gene was determined, and the deduced amino acid sequence is in accordance with the sequence of an internal peptide isolated from P39