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Cloning and sequencing of yajC and secD homologs of Brucella abortus and demonstration of immune responses to YajC in mice vaccinated with B. abortus RB51.
Infect Immun 1998; 66(12):5684-91II

Abstract

To identify Brucella antigens that are potentially involved in stimulating a protective cell-mediated immune response, a gene library of Brucella abortus 2308 was screened for the expression of antigens reacting with immunoglobulin G2a antibodies from BALB/c mice vaccinated with B. abortus RB51. One selected positive clone (clone MCB68) contained an insert of 2.6 kb; nucleotide sequence analysis of this insert revealed two open reading frames (ORFs). The deduced amino acid sequences of the first and second ORFs had significant similarities with the YajC and SecD proteins, respectively, of several bacterial species. Both the YajC and SecD proteins were expressed in Escherichia coli as fusion proteins with maltose binding protein (MBP). In Western blots, sera from mice vaccinated with B. abortus RB51 recognized YajC but not SecD. Further Western blot analysis with purified recombinant YajC protein indicated that mice inoculated with B. abortus 19 or 2308 or B. melitensis RM1 also produced antibodies to YajC. In response to in vitro stimulation with recombinant MBP-YajC fusion protein, splenocytes from mice vaccinated with B. abortus RB51 were able to proliferate and produce gamma interferon but not interleukin-4. This study demonstrates, for the first time, the involvement of YajC protein in an immune response to an infectious agent.

Authors+Show Affiliations

Center for Molecular Medicine and Infectious Diseases, Department of Biomedical Sciences and Pathobiology, VA-MD Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061-0342, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

9826342

Citation

Vemulapalli, R, et al. "Cloning and Sequencing of yajC and secD Homologs of Brucella Abortus and Demonstration of Immune Responses to YajC in Mice Vaccinated With B. Abortus RB51." Infection and Immunity, vol. 66, no. 12, 1998, pp. 5684-91.
Vemulapalli R, Duncan AJ, Boyle SM, et al. Cloning and sequencing of yajC and secD homologs of Brucella abortus and demonstration of immune responses to YajC in mice vaccinated with B. abortus RB51. Infect Immun. 1998;66(12):5684-91.
Vemulapalli, R., Duncan, A. J., Boyle, S. M., Sriranganathan, N., Toth, T. E., & Schurig, G. G. (1998). Cloning and sequencing of yajC and secD homologs of Brucella abortus and demonstration of immune responses to YajC in mice vaccinated with B. abortus RB51. Infection and Immunity, 66(12), pp. 5684-91.
Vemulapalli R, et al. Cloning and Sequencing of yajC and secD Homologs of Brucella Abortus and Demonstration of Immune Responses to YajC in Mice Vaccinated With B. Abortus RB51. Infect Immun. 1998;66(12):5684-91. PubMed PMID: 9826342.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Cloning and sequencing of yajC and secD homologs of Brucella abortus and demonstration of immune responses to YajC in mice vaccinated with B. abortus RB51. AU - Vemulapalli,R, AU - Duncan,A J, AU - Boyle,S M, AU - Sriranganathan,N, AU - Toth,T E, AU - Schurig,G G, PY - 1998/11/24/pubmed PY - 1998/11/24/medline PY - 1998/11/24/entrez SP - 5684 EP - 91 JF - Infection and immunity JO - Infect. Immun. VL - 66 IS - 12 N2 - To identify Brucella antigens that are potentially involved in stimulating a protective cell-mediated immune response, a gene library of Brucella abortus 2308 was screened for the expression of antigens reacting with immunoglobulin G2a antibodies from BALB/c mice vaccinated with B. abortus RB51. One selected positive clone (clone MCB68) contained an insert of 2.6 kb; nucleotide sequence analysis of this insert revealed two open reading frames (ORFs). The deduced amino acid sequences of the first and second ORFs had significant similarities with the YajC and SecD proteins, respectively, of several bacterial species. Both the YajC and SecD proteins were expressed in Escherichia coli as fusion proteins with maltose binding protein (MBP). In Western blots, sera from mice vaccinated with B. abortus RB51 recognized YajC but not SecD. Further Western blot analysis with purified recombinant YajC protein indicated that mice inoculated with B. abortus 19 or 2308 or B. melitensis RM1 also produced antibodies to YajC. In response to in vitro stimulation with recombinant MBP-YajC fusion protein, splenocytes from mice vaccinated with B. abortus RB51 were able to proliferate and produce gamma interferon but not interleukin-4. This study demonstrates, for the first time, the involvement of YajC protein in an immune response to an infectious agent. SN - 0019-9567 UR - https://www.unboundmedicine.com/medline/citation/9826342/Cloning_and_sequencing_of_yajC_and_secD_homologs_of_Brucella_abortus_and_demonstration_of_immune_responses_to_YajC_in_mice_vaccinated_with_B__abortus_RB51_ L2 - http://iai.asm.org/cgi/pmidlookup?view=long&pmid=9826342 DB - PRIME DP - Unbound Medicine ER -