A Productive Replication of Hyphantria cunea Nucleopolyhedrovirus in Lymantria dispar Cell Line

  • Demir, Ismail (Department of Biology, Faculty of Arts and Sciences, Karadeniz Technical University) ;
  • Demirbag, Zihni (Department of Biology, Faculty of Arts and Sciences, Karadeniz Technical University)
  • Published : 2006.10.31

Abstract

In this study, comparative replicational properties of Hyphantria cunea nucleopolyhedrovirus (HycuNPV) in Lymantria dispar (IPLB-LdElta) and Spodoptera frugiperda (IPLB-Sf21) cell lines were investigated. Our microscopic observations showed that cytopathic effects (CPEs) in LdElta cells appeared 12 h later than those in Sf21 cells. Whereas polyhedral inclusion bodies (PIBs) formed at 48 h postinfection (p.i.) in LdElta cells, it formed at 36 h p.i. in Sf21 cells. Extracellular virus production determined according to the 50% tissue culture infective dose ($TCID_{50}$) method in LdElta cells started about 12 h later when compared with Sf21 cells. Titers of extracellular virus in LdElta and Sf21 cells were calculated as $1.77{\times}10^9$ plaque forming units (PFU)/ml and $5.6{\times}10^9PFU/ml$, respectively, at 72 h p.i. We also showed that viral DNA replication began at 12 h p.i. in both cell lines. Viral protein synthesis was determined by SDS-polyacrylamide gel electrophoresis (PAGE) and polyhedrin synthesis was observed at 12 h p.i. in both cell lines. The results indicate that while the synthesis of macromolecules is 12 h later and production of extracellular virus is almost 3-fold lower in LdElta cells compared with those in Sf21 cells, the LdElta cell line is still a productive cell line for infection of HycuNPV.

Keywords

References

  1. Adams, J. R. and J. T. McClintock. 1991. Baculoviridae. Nuclear polyhedrosis viruses. Part 1. Nuclear polyhedrosis viruses of insects, pp. 89-204. In J. R. Adams and J. R. Bonami (eds.), Atlas of Invertebrate Viruses. CRC Press, Boca Raton
  2. Blissard, G. W. 1996. Baculovirus-insect cell interactions, pp. 73-93. In J. M. Vlak, C. D. De Gooijer, J. Tramper, and H. G. Miltenburger (eds.), Insect Cell Cultures: Fundamental and Applied Aspects. Kluwer Academic Publishers, Dordrecht, The Netherlands
  3. Bradford, M. B., G. W. Blissard, and G. F. Rohrmann. 1990. Characterization of the infection cycle of the Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus in Lymantria dispar cells. J. Gen. Virol. 71: 2841-2846 https://doi.org/10.1099/0022-1317-71-12-2841
  4. Brown, M. and P. Faulkner. 1975. Factors affecting the yield of virus in a cloned cell line of Trichoplusia ni infected with a nuclear polyhedrosis virus. J. Invertebr. Pathol. 26: 251-257 https://doi.org/10.1016/0022-2011(75)90057-9
  5. Castro, M. E. B., M. L. Souza, S. Arauja, and S. L. Bilimoria. 1997. Replication of Anticarsia gemmatalis nuclear polyhedrosis virus four lepidopteran cell lines. J. Invertebr. Pathol. 69: 40-45 https://doi.org/10.1006/jipa.1996.4624
  6. Choe, Y. K., S. M. Byun, H. H. Lee, T. W. Chung, H. Y. Park, and S. K. Kang. 1986. A nuclear polyhedrosis virus of Hyphantria cunea replicates in vitro. Biotechnol. Lett. 8: 853-858 https://doi.org/10.1007/BF01078644
  7. Demir, I., R. Nalcacioglu, Z. Demirbag, A. O. Kilic, and A. O. Belduz. 2000. Expression of cryIVA and cryIVD genes of Bacillus thuringiensis in baculovirus expression system. IOBC/WPAS wprs Bull. 23: 267-274
  8. Demirbag, Z. 1993. Comparative replication of Autographa californica nuclear polyhedrosis virus in abortive and productive infections of insect cell lines. In PhD Thesis. Texas Tech. University, Texas, U.S.A
  9. Demirbag, Z., A. O. Belduz, O. Erturk, and I. Demir. 1997. Comparative replication of Autographa californica nuclear polyhedrosis virus in insect cell cultures. Tr. J. Biol. 21: 63-70
  10. De Moreas, R. R. and J. E. Maruniak. 1997. Detection and identification of multiple baculovirus using the polymerase chain reaction (PCR) and restriction endonuclease analysis. J. Virol. Methods 63: 209-217 https://doi.org/10.1016/S0166-0934(96)02130-1
  11. Hostetter, D. L. and B. Puttler. 1991. A new broad host spectrum nuclear polyhedrosis virus isolated from a celery looper, Anagrapha falcifera (Kirby) (Lepidoptera: Noctuidae). Environ. Entomol. 20: 1481-1488
  12. Huber, J. 1986. Use of baculoviruses in pest management programs, pp. 181-202. In R. R. Granados and B. A. Federici (eds.), The Biology of Baculoviruses, Vol. 2. CRC Press, Boca Raton, Florida
  13. Invitrogen. 1996. MaxBac 2.0 Transfection and expression manual, pp. 35-36. In Invitrogen Corporation, A Manual of Methods for Baculovirus Expression. San Diego, U.S.A
  14. Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685 https://doi.org/10.1038/227680a0
  15. Lavina-Caoili, B. A., K. Kamiya, S. Kawamura, M. Ikeda, and M. Kobayashi. 2001. Comparative in vitro analysis of geographic variants of nucleopolyhedrovirus of Spodoptera litura isolated from China and The Philippines. J. Insect Biotechnol. Sericol. 70: 199-209
  16. Lee, H. H. 1987. Replication and cloning of Hyphantria cunea nuclear polyhedrosis virus in Spodoptera frugiperda cell line. HG. J. Gen. Eng. 2: 3-7
  17. Lee, H. H. and K. K. Lee. 1988. Isolation, complementation and partial characterization of temperature-sensitive mutants of the baculovirus Hyphantria cunea nuclear polyhedrosis virus. J. Gen. Virol. 69: 1299-1306 https://doi.org/10.1099/0022-1317-69-6-1299
  18. Levin, D. B. and J. Huang. 1999. Spodoptera littoralis type B nucleopolyhedrovirus infection of a grasshopper cell line. J. Invertebr. Pathol. 74: 184-192 https://doi.org/10.1006/jipa.1999.4874
  19. Liu, H. S. and S. L. Bilimoria. 1990. Infected cell specific protein and viral DNA synthesis in productive and abortive infections of Spodoptera frugiperda nuclear polyhedrosis virus. Arch. Virol. 115: 101-113 https://doi.org/10.1007/BF01310626
  20. Lu, A. and L. K. Miller. 1997. Regulation of baculovirus late and very late gene expression, pp. 193-216. In L. K. Miller (ed.), The Baculoviruses. Plenum Press, New York
  21. Lynn, D. E. 2003. Comparative susceptibilities of twelve cell lines to infection by three baculoviruses. J. Invertebr. Pathol. 82: 129-131 https://doi.org/10.1016/S0022-2011(03)00002-8
  22. Lynn, D. E. 2003. Comparative susceptibilities of insect cell lines to infection by the occlusion-body derived phenotype of baculoviruses. J. Invertebr. Pathol. 83: 215-222 https://doi.org/10.1016/S0022-2011(03)00091-0
  23. Lynn, D. E., E. M. Dougherty, J. T. McClintock, and M. Loeb. 1988. Development of cell lines from various tissues of Lepidoptera, pp. 239-242. In Y. Kuroda, E. Kurstack and K. Maramorosch (eds.), Invertabrate and Fish Tissues Culture. Japan Scientific Society Press, Tokyo/Springer-Verlag, Berlin
  24. McClintock, J. T., E. M. Dougherty, and R. M. Wiener. 1986. Semipermissive replication of a nuclear polyhedrosis virus of Autographa californica in a gypsy moth cell line. J. Virol. 57: 197-204
  25. O'Reilly, D. R., L. K. Miller, and V. A. Luckow. 1992. Baculovirus Expression Vectors: A Laboratory Manual. New York, Freeman, W.H. and Company
  26. Reed, L. and H. Muench. 1938. A simple method for estimating fifty percent end points. Am. J. Hyg. 27: 493-497
  27. Riegel, C. I. and J. M. Slavicek. 1997. Characterization of the replication cycle of the Lymantria dispar nuclear polyhedrosis virus. Virus Res. 51: 9-17 https://doi.org/10.1016/S0168-1702(97)00075-0
  28. Sambrook, J., E. F. Fritsch, and T. Maniatis. 1989. Molecular Cloning, 2nd Ed. Cold Spring Habor Laboratory Press, New York
  29. Shirata, N., M. Ikeda, K. Kamiya, S. Kawamura, Y. Kunimi, and M. Kobayashi. 1999. Replication of nucleopolyhedrosisviruses of Autographa californica (Lepidoptera: Noctuidae), Bombyx mori (Lepidoptera: Bombycidae), Hyphantria cunea (Lepidoptera: Arctiidae) and Spodoptera exigua (Lepidoptera: Noctuidae) in four lepidopteran cell lines. Appl. Entomol. Zool. 34: 507-516 https://doi.org/10.1303/aez.34.507
  30. Vaughn, J. L., R. M. Goodman, G. T. Tomkins, and P. McCawley. 1977. The establishment of two cell lines from the insect Spodoptera frugiperda (Lepidoptera, Noctiudae). In Vitro 13: 213-217 https://doi.org/10.1007/BF02615077
  31. Volkman, L. E., G. W. Blissard, P. Friesen, B. A. Keddie, R. Possee, and D. A. Theilmann. 1995. Baculoviridae, pp. 104-111. In F. A. Murphy, C. M. Fauquet, D. H. L. Bishop, S. A. Ghabrial, A. W. Jarvis, G. P. Martelli, M. A. Mayo, and M. D. Summers (eds.), Virus Taxonomy, Sixth Report of the International Committee on Taxonomy of Viruses. Springer-Verlag, New York
  32. Williams, G. V. and P. Faulkner. 1996. Replication patterns and cytopathology of cells infected with baculoviruses, pp. 95-110. In J. M. Vlak, C. D. De Gooijer, J. Tramper, and H. G. Miltenburger (eds.), Insect Cell Cultures: Fundamental and Applied Aspects. Kluwer Academic Publishers, Dordrecht, The Netherlands
  33. Woo, S. D. 2001. Rapid detection of multiple nucleopolyhedroviruses using polymerase chain reaction. Mol. Cells 11: 334-340
  34. Wu, F. Q., B. A. Lavina, M. Ikeda, N. Shirata, Y. X. Cai, S. X. Pan, and M. Kobayashi. 2000. Cloning and biological characterization of Spodoptera exigua nucleopolyhedroviruses isolated in China. J. Seric. Sci. Jpn. 69: 177-189