IBDV에 대한 단크론항체 생산 및 진단적 응용
Production and diagnostic application of monoclonal antibodies against infectious bursal disease virus
1
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Infectious bursal disease (IBD) caused by infectious bursal disease virus (IBDV) is a highly contagious viral disease in chicken. It causes heavy economic loss by immune suppression and high mortality. The IBDV, designated Avibirnavirus in the Family Birnaviridae, has a double-stranded RNA genome formed by two segments, segment A and segment B. Segment A encodes a 108 KDa polypeptide that is self-cleaved to produce pVP2, VP3 and VP4, and later pVP2 is cleaved to VP2. The VP2 contains the antigenic regions responsible for elicitation of neutralizing antibodies and VP3 is a major immunogenic protein of IBDV. In this study, monoclonal antibodies (MAbs) specific for IBDV were produced and characterized. All 15 MAbs were specific for IBDV and did not react with other viruses used in this study. The protein specificity of MAbs was determined by comparing the reactivity patterns of each MAb with IBDV VP2 and VP234 recombinant baculoviruses and Western blot analysis. As a result,7 MAbs (1F5, 2C8, 2F4, 3C7, 4C3, 6F11, 6G5) and 5 MAbs (2A4, 2G2, 3F5, 3G2, 4F10) were specific for VP2 and VP3, respectively. The protein specificity of 3 MAbs (2B8, 3F7, 3F8) were not determined. Five (2C8, 2F4, 4C3, 6F11, 6G5) of the VP2-specific MAbs had a neutralizing activity against IBDV. Some MAbs reacted with IBDV-infected bursa of Fabricius by indirect fluorescence antibody (IFA) and immunohistochemistry (IHC) assay. The MAbs produced in this study would be used for diagnostic reagents for the detection of IBDV infection.
Abstract
Infectious bursal disease (IBD) caused by infectious bursal disease virus (IBDV) is a highly contagious viral disease in chicken. It causes heavy economic loss by immune suppression and high mortality. The IBDV, designated Avibirnavirus in the Family Birnaviridae, has a double-stranded RNA genome formed by two segments, segment A and segment B. Segment A encodes a 108 KDa polypeptide that is self-cleaved to produce pVP2, VP3 and VP4, and later pVP2 is cleaved to VP2. The VP2 contains the antigenic regions responsible for elicitation of neutralizing antibodies and VP3 is a major immunogenic protein of IBDV. In this study, monoclonal antibodies (MAbs) specific for IBDV were produced and characterized. All 15 MAbs were specific for IBDV and did not react with other viruses used in this study. The protein specificity of MAbs was determined by comparing the reactivity patterns of each MAb with IBDV VP2 and VP234 recombinant baculoviruses and Western blot analysis. As a result,7 MAbs (1F5, 2C8, 2F4, 3C7, 4C3, 6F11, 6G5) and 5 MAbs (2A4, 2G2, 3F5, 3G2, 4F10) were specific for VP2 and VP3, respectively. The protein specificity of 3 MAbs (2B8, 3F7, 3F8) were not determined. Five (2C8, 2F4, 4C3, 6F11, 6G5) of the VP2-specific MAbs had a neutralizing activity against IBDV. Some MAbs reacted with IBDV-infected bursa of Fabricius by indirect fluorescence antibody (IFA) and immunohistochemistry (IHC) assay. The MAbs produced in this study would be used for diagnostic reagents for the detection of IBDV infection.
참고문헌 (26)
- 權容國, "傳染性 F囊病 바이러스 國內 分離株 SH", 92 病原性硏究, 1995, pp.37, 637~647
- 이영옥, "국내종계의 infectious bursal disease virus의 감염상황 및 분리주의 생물학적 특성", 농사시험보고서(축산, 가축위생편), 23, pp.136~142 (1981)
- Bayliss CD, "A comparison of the sequences of segment A of four infectious bursal disease virus strains and identification of a variable region in VP2", J Gen Virol, 71(6), pp.1303~1312 (1990)
- Becht H, "Comparative studies on structural and antigenic properties of two serotypes of infectious bursal disease virus", J Gen Virol, 69(3), pp.631~640 (1988)
- Berg TP, "Acute infectious bursal disease in poultry: a review", Avian Pathol, 29(3), pp.175~194 (2000)
- 최강석, "Rapid Detection of Infectious Bursal Disease Virus (IBDV) in Chickens by an Immunochromatographic Assay Kit", 한국가금학회지, 37(2), pp.167~171 (2010), 한국가금학회
- Coyle PV, "A simple standardised protocol for the production of monoclonal antibodies against viral and bacterial antigens", J Immunol Methods, 153(1), pp.81~84 (1992)
- Delmas B, "In Virus Taxonomy. Eighth r Iort of the International Committee on Taxonomy of Viruses: Birnaviridae", pp.561~569 (2004), Academic Press
- Fahey KJ, "Antibody to the 32K structural protein of infectious bursal disease virus neutralizes viral infectivity in vitro and confers protection on young chickens", J Gen Virol, 66(12), pp.2693~2702 (1985)
- Fahey KJ, "A conformational immunogen on VP-2 of infectious bursal disease virus that induces virus-neutralizing antibodies that passively protect chickens", J Gen Virol, 70(6), pp.1473~1481 (1989)
- Heine HG, "Sequence analysis and expression of the host-protective immunogen VP2 of a variant strain of infectious bursal disease virus which can circumvent vaccination with standard type I strains", J Gen Virol, 72(8), pp.1835~1843 (1991)
- Hudson PJ, "Genomic structure of the large RNA segment of infectious bursal disease virus", Nucleic Acids Res, 14(12), pp.5001~5012 (1986)
- Ismail NM, "Lack of pathogenicity of five serotype 2 infectious bursal disease viruses in chickens", Avian Dis, 32(4), pp.757~759 (1988)
- Woo-Jin Jeon, "Detection of Infectious Bursal Disease Virus by Double Antibody Sandwich ELISA", Journal of Bacteriology and Virology, 38(3), pp.139~147 (2008), 대한미생물학회
- Kang SY, "Reactivity of VP4-specific monoclonal antibodies to a serotype 4 porcine rotavirus with distinct serotypes of human(symptomatic and asymptomatic) and animal rotaviruses", J Clin Microbiol, 27(12), pp.2744~2750 (1989)
- Kibenge FS, "Growth of serotypes I and II and variant strains of infectious bursal disease virus in Vero cells", Avian Dis, 32(2), pp.298~303 (1988)
- Lim BL, "Adaptation of very virulent infectious bursal disease virus to chicken embryonic fibroblasts by site-directed mutagenesis of residues 279 and 284 of viral coat protein VP2", J Virol, 73(4), pp.2854~2862 (1999)
- Lukert PD, "Infectious bursal disease virus: growth and characterization in cell cultures", Avian Dis, 18(2), pp.243~250 (1974)
- Martinez-Torrecuadrada JL, "Antigenic properties and diagnostic potential of baculovirus-expressed infectious bursal disease virus proteins VPX and VP3", Clin Diagn Lab Immunol, 7(4), pp.645~651 (2000)
- Miller LK, "Baculoviruses for foreign gene expression in insect cells. Review", Biotechnology, 10, pp.457~465 (1988)
- Morser MJ, "Virus-specified polypeptides in cells infected with Semliki forest virus", J Gen Virol, 21(1), pp.19~29 (1973)
- Mundt E, "Identification of a novel viral protein in infectious bursal disease virus-infected cells", J Gen Virol, 76(2), pp.437~443 (1995)
- Pitcovski J, "Development and large-scale use of recombinant VP2 vaccine for the prevention of infectious bursal disease of chickens", Vaccine, 21(32), pp.4736~4743 (2003)
- Spies U, "Properties of RNA polymerase activity associated with infectious bursal disease virus and characterization of its reaction products", Virus Res, 8(2), pp.127~140 (1987)
- Towbin H, "Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications", Proc Natl Acad Sci USA, 76(9), pp.4350~4354 (1979)
- Vakharia VN, "Active and passive protection against variant and classic infectious bursal disease virus strains induced by baculovirus-expressed structural proteins", Vaccine, 12(5), pp.452~456 (1994)