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VETERINARY IMMUNOLOGY

Antigenic and Immunogenic Investigation of B-Cell Epitopes in the Nucleocapsid Protein of Peste des Petits Ruminants Virus

Kang-Seuk Choi, Jin-Ju Nah, Young-Joon Ko, Shien-Young Kang, Kyoung-Jin Yoon, Nam-In Jo
Kang-Seuk Choi
1Foreign Animal Disease Division, National Veterinary Research and Quarantine Service, Anyang, Kyoung-gi
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  • For correspondence: choiks@nvrqs.go.kr
Jin-Ju Nah
1Foreign Animal Disease Division, National Veterinary Research and Quarantine Service, Anyang, Kyoung-gi
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Young-Joon Ko
1Foreign Animal Disease Division, National Veterinary Research and Quarantine Service, Anyang, Kyoung-gi
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Shien-Young Kang
2College of Veterinary Medicine, Chungbuk National University, Heungduk-Ku, Cheongju, Chungbuk, Korea
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Kyoung-Jin Yoon
3Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, Iowa
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Nam-In Jo
1Foreign Animal Disease Division, National Veterinary Research and Quarantine Service, Anyang, Kyoung-gi
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DOI: 10.1128/CDLI.12.1.114-121.2005
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  • FIG. 1.
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    FIG. 1.

    Schematic diagram (A) of GST fusion protein constructs containing full-length or truncated nucleocapsid fragments of PPRV and Western immunoblot confirmation (B) of E. coli-expressed products using goat anti-PPRV serum.

  • FIG. 2.
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    FIG. 2.

    Reactivity of anti-PPRV nucleocapsid MAbs with native (A), baculovirus-expressed recombinant (B), or E. coli-expressed recombinant GST fusion nucleocapsid proteins (C) in an indirect ELISA.

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    FIG. 3.

    Mapping of MAb epitopes using GST fusion proteins containing partially overlapped fragments of PPRV nucleocapsid protein. GST-N1-525 (B), GST-N1-262 (C), GST-N1-447 (D), and GST-N405-521 (E) were separated electrophoretically and then transferred to nitrocellulose membranes. Baculovirus-expressed N protein rPPRV-N (A) was used as control. The binding of each MAb to GST fusion proteins on the membranes was detected using Western immunoblotting.

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    FIG. 4.

    Immunogenicity of seven epitopes on the nucleocapsid of PPRV as determined by the degree of blocking of their binding to the baculovirus-expressed recombinant PPRV N protein by anti-PPRV caprine sera (PPR 2-65, PPR 2-66, PPR 2-69, PPR 2-73, and PPR 2-81), PPRV hyperimmune serum (RPV positive), or anti-PRV bovine sera (RP K9061 and K9062) at dilutions of 1:20 (A) and 1:400 (B). Each epitope is designated by the name of each detector MAb.

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    FIG. 5.

    Average humoral antibody response to four immunodominant epitopes (MAbs 38-4, P-3H12, P-13A9, and P-11A6) of PPRV N protein in three goats experimentally infected with PPRV of lineage I as measured by a competitive ELISA.

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  • TABLE 1.

    Mapping of epitopes on the N protein of PPRV Nig75/1 straina

    Competing MAb (isotype)DomainLabeled MAb
    33-4P-3H12P-13A938-4P-14C6P-9H10P-11A6
    33-4 (IgG1)A-I++−−−−−−
    P-3H12 (IgG2a)A-II−+++++−−−−
    P-13A9 (IgG2b)A-II−+++++++−−−
    38-4 (IgG1)A-II−+++++++++−−−
    P-14C6 (IgM)C-I−++++−++−−
    P-9H10 (IgG1)C-II−++−−+++−
    P-11A6 (IgG2b)C-II−++++++−−++++++
    • ↵ a −, <50% competition by unlabeled MAb at 1:100; +, >50% competition by unlabeled MAb at 1:100; ++, >50% competition by unlabeled MAb at 1:1,000; +++, >50% competition by unlabeled MAb at 1:10,000.

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Antigenic and Immunogenic Investigation of B-Cell Epitopes in the Nucleocapsid Protein of Peste des Petits Ruminants Virus
Kang-Seuk Choi, Jin-Ju Nah, Young-Joon Ko, Shien-Young Kang, Kyoung-Jin Yoon, Nam-In Jo
Clinical and Diagnostic Laboratory Immunology Jan 2005, 12 (1) 114-121; DOI: 10.1128/CDLI.12.1.114-121.2005

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Antigenic and Immunogenic Investigation of B-Cell Epitopes in the Nucleocapsid Protein of Peste des Petits Ruminants Virus
Kang-Seuk Choi, Jin-Ju Nah, Young-Joon Ko, Shien-Young Kang, Kyoung-Jin Yoon, Nam-In Jo
Clinical and Diagnostic Laboratory Immunology Jan 2005, 12 (1) 114-121; DOI: 10.1128/CDLI.12.1.114-121.2005
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