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Vero 세포적응 뉴캣슬병 바이러스의 생물학적 특성

Biological Properties of Vero Cell-Adapted Newcastle Disease Virus

  • 최강석 (농림수산검역검사본부 조류질병과) ;
  • 박미자 (농림수산검역검사본부 조류질병과) ;
  • 계수정 (농림수산검역검사본부 조류질병과) ;
  • 김지예 (농림수산검역검사본부 조류질병과) ;
  • 권준헌 (농림수산검역검사본부 조류질병과)
  • Choi, Kang-Seuk (Avian Disease Division, Animal, Plant and Fisheries Quarantine and Inspection Agency) ;
  • Park, Mi-Ja (Avian Disease Division, Animal, Plant and Fisheries Quarantine and Inspection Agency) ;
  • Kye, Soo-Jeong (Avian Disease Division, Animal, Plant and Fisheries Quarantine and Inspection Agency) ;
  • Kim, Ji-Ye (Avian Disease Division, Animal, Plant and Fisheries Quarantine and Inspection Agency) ;
  • Kwon, Jun-Hun (Avian Disease Division, Animal, Plant and Fisheries Quarantine and Inspection Agency)
  • 투고 : 2012.03.10
  • 심사 : 2012.05.16
  • 발행 : 2012.06.30

초록

본 연구를 통하여 Vero 세포에서 강독형 NDV인 Kr005주를 55대 연속 계대하여 Vero 세포에 적응된 Kr005/V 주를 작성하였다. Kr005/V 주는 Vero 세포에서 $10^{7.8}/mL$의 높은 바이러스 증식성을 보였으며, 감염세포는 cell rounding과 같은 CPE를 형성하였다. 또한 이열성(heat labile) NDV Kr005주와 달리 NDV Kr005/V 주는 내열성(thermostable)을 보였다. 계태아에서 평균 치사 시간(MDT)를 측정한 결과, 강독형인 Kr005주(49.6시간의 MDT)와 달리 Kr005/V 주는 120시간이상의 MDT를 보여 약독형 바이러스(lentogenic virus)로 분류되었다. Kr005/V 주의 HN 단백질과 F 단백질의 염기서열을 분석한 결과, HN 단백질의 443번째 아미노산에서만 변이(T에서 S)가 관찰되었다. Kr005/V 주와 Vero 세포를 사용하여 야외 닭 혈청을 대상으로 혈청중화 항체가를 측정한 결과, Kr005주와 CEF 세포를 사용하여 측정한 혈청중화 항체가와 97%의 상관성을 나타내었다.

Newcastle disease virus (NDV) Kr005/V strain was generated through 55 serial passages of NDV Kr005 strain in Vero cells. The Kr005/V virus yielded high infective titers of $10^{7.8}$ $TCID_{50}/mL$ in Vero cells and the infected cells showed cytopathic effects such as marked cell rounding, though less frequent syncytia. The Kr005/V virus was heat-stable and classified into the lentogenic type with a Mean Death Time (MDT) of 120h or greater while the Kr005 strain was heat-labile and velogenic (MDT of 49.6 h). Only the single amino acid substitution (T to S) was observed at position 433 of the HN protein of the Kr005/V strain, whereas no amino acid change was found in the F protein. The Kr005/V input virus correlated well (correlation coefficient $r^2$=0.97) with the Kr005 virus when ten field sera were tested by virus neutralization test. The biological properties and usefulness of Vero cell-adapted Kr005/V virus were discussed.

키워드

참고문헌

  1. Ahamed T, Hossain KM, Billah MM, Islam KMD, Ahasan MM, Islam ME 2004 Adaptation of Newcastle disease virus (NDV) on Vero cell line. Int J Poult Sci 3: 153-156. https://doi.org/10.3923/ijps.2004.153.156
  2. Alexander DJ 1988 Newcastle Disease Diagnosis. pp 147-160. In: Newcastle Disease. Kluwer Academic Publishers, Boston.
  3. Alexander DJ 2003 Newcastle Disease. pp 64-87 In: Diseases of Poultry, 11th edition. Iowa State University Press, Ames, IA.
  4. Bankowski RA 1964 Propagation of Newcastle Disease Virus. pp 231-246. In: Newcastle Disease Virus: An Evolving Pathogen. University Wisconsin Press, Madison.
  5. de Leeuw OS, Koch G, Hartog L, Ravenshorst N, Peeters BP 2005 Virulence of Newcastle disease virus is determined by the cleavage site of the fusion protein and by both the stem region and globular head of the haemagglutinin-neuraminidase protein. J Gen Virol 86:1759-1769. https://doi.org/10.1099/vir.0.80822-0
  6. Fenner F, McAuslan BB, Mims CA, Sambrook J, White DO 1974 The Biology of Animal Viruses. p 40. Academic Press, London.
  7. Gravel KA, Morrison TG 2003 Interacting domains of the HN and F proteins of Newcastle disease virus. J Virol 77:11040-11049. https://doi.org/10.1128/JVI.77.20.11040-11049.2003
  8. Heminway BR, Yu Y, Galinski MS 1994 Paramyxovirus mediated cell fusion requires co-expression of both the fusion and hemagglutinin-neuraminidase glycoproteins. Virus Res 31:1-16. https://doi.org/10.1016/0168-1702(94)90066-3
  9. Horvath CM, Paterson RG, Shaughnessy MA, Wood R, Lamb RA 1992 Biological activity of paramyxovirus fusion proteins: factors influencing formation of syncytia. J Virol 66:4564-4569.
  10. Huang Z, Panda A, Elankumaran S, Govindarajan D, Rockemann DD, Samal SK 2004 The hemagglutinin-neuraminidase protein of Newcastle disease virus determines tropism and virulence. J Virol 78:4176-4184. https://doi.org/10.1128/JVI.78.8.4176-4184.2004
  11. Iorio RM, Glickman RL 1992 Fusion mutants of Newcastle disease virus selected with monoclonal antibodies to the hemagglutinin-neuraminidase. J Virol 66:6626-6633.
  12. Khattar SK, Yan Y, Panda A, Collins PL, Samal SK 2009 A Y526Q mutation in the Newcastle disease virus HN protein reduces its functional activities and attenuates virus replication and pathogenicity. J virol 83:7779-82. https://doi.org/10.1128/JVI.00536-09
  13. Kumanan K, Venkatesan RA 1994 Adaptation of Newcastle disease virus strains to different cell culture. Indian J Comp Microbiol Immunol Infect Dis 15:39-41.
  14. Kumanan K, Vijayarani K, Padmanabhan VD 1993 Adaptation of a velogenic Newcastle disease virus isolate to Vero cells. Indian Vet J 70:993-995.
  15. Lee YJ, Sung HW, Choi JG, Kim JH, Song CS 2004 Molecular epidemiology of Newcastle disease viruses isolated in South Korea using sequencing of the fusion protein cleavage site region and phylogenetic relationships. Avian Pathol 33:482-491. https://doi.org/10.1080/03079450400003700
  16. Lee YJ, Sung HW, Choi JG, Lee EK, Yoon Y, Kim JH, Song CS 2008 Protection of chickens from Newcastle disease with a recombinant baculovirus subunit vaccine expressing the fusion and hemagglutinin-neuraminidase proteins. J Vet Sci 9:301-308. https://doi.org/10.4142/jvs.2008.9.3.301
  17. Lomniczi B 1975 Thermostability of Newcastle disease virus strains of different virulence. Arch Virol 47:249-255. https://doi.org/10.1007/BF01317812
  18. Mayo MA 2002 A summary of taxonomic changes recently approved by ICTV. Arch Virol 147:1655-1663. https://doi.org/10.1007/s007050200039
  19. Melanson VR, Iorio RM 2004 Amino acid substitutions in the F-specific domain in the stalk of the Newcastle disease virus HN protein modulate fusion and interfere with its interaction with the F protein. J Virol 78:13053-13061. https://doi.org/10.1128/JVI.78.23.13053-13061.2004
  20. Mohan CM, Dey S, Kumanan K, Manohar BM, Nainar AM 2007 Adaptation of a velogenic Newcastle disease virus to Vero cells: assessing the molecular changes before and after adaptation. Vet Res Commun 31:371-383. https://doi.org/10.1007/s11259-006-3502-2
  21. Panda A, Huang Z, Elankumaran S, Rockemann DD, Samal SK 2004 Role of 250 fusion protein cleavage site in the virulence of Newcastle disease virus. Microb Pathog 36:1-10. https://doi.org/10.1016/j.micpath.2003.07.003
  22. Peeters BP, de Leeuw OS, Koch G, Gielkens AL 1999 Rescue of Newcastle 253 disease virus from cloned cDNA: evidence that cleavability of the fusion protein is a 254 major determinant for virulence. J Virol 73:5001-5009.
  23. Reddy GS, Srinivasan VA 1992 Use of BHK cell culture-adapted Newcastle disease virus for immunization of chicks. Vaccine 10:164-166. https://doi.org/10.1016/0264-410X(92)90006-6
  24. Reed LJ, Muench H 1938 A simple method for estimating fifty percent endpoints. Am J Hyg 27:493-497.
  25. Romer-Oberdörfer A, Werner O, Veits J, Mebatsion T, Mettenleiter TC 2003 Contribution of the length of the HN protein and the sequence of the F protein cleavage site to Newcastle disease virus pathogenicity. J Gen Virol 84:3121-3129. https://doi.org/10.1099/vir.0.19416-0
  26. Takimoto T, Taylor GL, Connaris HC, Crennell SJ, Portner A 2002 Role of the hemagglutinin-neuraminidase protein in the mechanism of paramyxovirus-cell membrane fusion. J Virol 76:13028-13033. https://doi.org/10.1128/JVI.76.24.13028-13033.2002
  27. 김재홍 송창선 1992 최근의 닭 뉴캣슬병 만연에 따른 원인과 바이러스의 성장, 진단 및 백신 운용에 관한 고찰. 한국가금학회지 19:65-76.
  28. 박근식 1979 한국에 있어서 뉴캣슬병 발생의 역학적 조사. 한국가금학회지 6:38-46.
  29. 최강석 2010 최근 국내에서 유행하는 뉴캣슬병 바이러스의 특성 고찰. 한국가금학회지 37:89-99.