Korean Journal of Veterinary Service (한국동물위생학회지)

The Korean Society of Veterinary Service (한국동물위생학회)
권호 표지

Production and characterization of monoclonal antibodies against rabies virus

광견병바이러스에 대한 단크론항체 생산 및 특성

  • Received : 2010.04.09
  • Accepted : 2010.06.11
  • Published : 2010.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

Rabies virus which belongs to the genus Lyssavirus of the family Rhabdoviridae is known as a highly neurotropic virus and causes fatal encephalitis accompanied by severe neurological symptoms in almost all mammals, including humans. In this study, monoclonal antibodies (MAbs) against rabies virus were produced, characterized and applications of MAbs as diagnostic reagents were assessed Spleen and inguinal lymph node cells from Balb/c mouse immunized with purified rabies virus were fused with SP2/O myeloma cells using polyethylene glycol (PEG) and hybridoma cells producing rabies virus-specific MAbs were screened by an indirect fluorescent antibody test. A total of ten MAbs were produced against rabies virus. The protein specificity and neutralizing activity of MAbs were determined by Western blot analysis and fluorescent antibody virus neutralization test, respectively. As a result, two MAbs, 5G3 and 6H4 had specificity for nucleoprotein (N protein) and two other MAbs, 5B1 and 5C1 had neutralizing activity for rabies virus. Some MAbs recognized the rabies virus-infected bovine brain stem cells by immunohistochemistry (IHC) assay. In conclusion, it was confirmed that MAbs produced in this study were rabies virusspecific and could be used as reliable diagnostic reagents for the detection of rabies virus.

Keywords

References

  1. Aaslestad HG, Wiktor TJ. 1972. Recovery of protective activity in rabies virus vaccines concentrated and purified by four different methods. Appl Microbiol 24: 37-43.
  2. Badrane H, Tordo N. 2001. Host switching in Iyssavirus history from the Chiroptera to the Carnivora orders. J Virol 75: 8096-8104. https://doi.org/10.1128/JVI.75.17.8096-8104.2001
  3. Beran GW, Steele JH. 1994. Handbook of Zoonoses, Section B. CRC Press. Boca Raton: 307-308.
  4. Blancou J. 1988. Ecology and epidemiology of fox rabies. Rev Infect Dis 10: 606-609. https://doi.org/10.1093/clinids/10.Supplement_4.S606
  5. Brochier B, Kieny MP, Costy F, Coppens P, Bauduin B, Lecocq JP, Languet B, Chappuis G, Desmettre P, Afiademanyo K, Libois R, Pastoret PP. 1991. Largescale eradication of rabies using recombinant vacciniarabies vaccine. Nature 354: 520-522. https://doi.org/10.1038/354520a0
  6. Coyle PV, Wyatt D, McCaughey C, O'Neill HJ. 1992. A simple standardised protocol for the production of monoclonal antibodies against viral and bacterial antigens. J Immunol Methods 153: 81-84. https://doi.org/10.1016/0022-1759(92)90308-G
  7. Cox JH, Dietzschold B, Schneider LG. 1977. Rabies virus glycoprotein. II. Biological and serological characterization. Infect Immun 16: 754-759.
  8. Dietzschold B, Wang HH, Rupprecht CE, Celis E, Tollis M, Ertl H, Heber-Katz E, Koprowski H. 1987. Induction of protective immunity against rabies by immunization with rabies virus ribonucleoprotein. Proc Natl Acad Sci USA 84: 9165-9169. https://doi.org/10.1073/pnas.84.24.9165
  9. Hyun BH, Lee KK, Kim IJ, Lee KW, Park HJ, Lee OS, An SH, Lee JB. 2005. Molecular epidemiology of rabies virus isolates from South Korea. Virus Res 114: 113-125. https://doi.org/10.1016/j.virusres.2005.06.004
  10. Kang SY, Saif LJ, Miller KL. 1989. 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: 2744-2750.
  11. Kohler G, Milstein C. 1975. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256: 495-497. https://doi.org/10.1038/256495a0
  12. Lahaye X, Vidy A, Pomier C, Obiang L, Harper F, Gaudin Y, Blondel D. 2009. Functional characterization of Negri bodies (NBs) in rabies virus-infected cells: Evidence that NBs are sites of viral transcription and replication. J Virol 83: 7948-7958. https://doi.org/10.1128/JVI.00554-09
  13. Lee JH, Lee MJ, Lee JB, Kim JS, Bae CS, Lee WC. 2001. Review of canine rabies prevalence under two diffe-rent vaccination programmes in Korea. Vet Rec 148: 511-512. https://doi.org/10.1136/vr.148.16.511
  14. Lodmell DL, Esposito JJ, Ewalt LC. 1993. Rabies virus antinucleoprotein antibody protects against rabies virus challenge in vivo and inhibits rabies virus replication in vitro. J Virol 67: 6080-6086.
  15. Orlik O, Altaner C. 1988. Modification of hybridoma technology which improve the yield of monoclonal antibody producing cells. J Immunol Methods 115: 55-59. https://doi.org/10.1016/0022-1759(88)90309-2
  16. Towbin H, Staehelin T, Gordon J. 1985. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc NatI Acad Sci USA 76: 4350-4354.
  17. World Health Organization. 1998. World survey of rabies no. 32 for the year 1996. WHO document EMC/ZDI/98.4. World Health Organization, Geneva, Switzerland.
  18. Wunner WH, Larson JK, Dietzschold B, Smith CL. 1988. The molecular biology of rabies viruses. Rev Infect Dis 10: S771-S784. https://doi.org/10.1093/clinids/10.Supplement_4.S771