İsmail DEMİR, Nurten GÜREL, Remziye NALÇACIOĞLU, Zihni DEMİRBAĞ

Comparative Susceptibilities of Six Insect Cell Lines to Infection by Malacosoma neustria Nucleopolyhedrovirus (ManeNPV)

In this study, 6 lepidopteran cell lines derived from Trichoplusia ni (Tn5), Lymantria dispar (LdElta), Bombyx mori (Bm-5), Spodoptera frugiperda (Sf21), Spodoptera exigua (Se-1), and Choristoneura fumiferana (Cf-124T) were investigated for susceptibility to Malacosoma neustria nucleopolyhedrovirus (ManeNPV) isolated in Turkey. Infection of Malacosoma disstria (Md203) cell line by ManeNPV was used as a control. In ManeNPV-Md203 infection, polyhedral inclusion bodies (PIBs) were detected at 36 h p.i. and observed in all infected cells by 120 h p.i., and the infectious virus titer increased 103-fold. At 24 h p.i., the efficient replication of viral DNA was detected. In this study, we determined that Tn5 and LdElta cells were productive to ManeNPV, and the replication of viral DNA occurred at 24 h p.i. PIB productions were determined in both cell lines at 36 h p.i., and the infectious virus titer in both cell lines increased 10-fold. By contrast, infections of Sf21 cells were not found productive. However, infected cells had apoptosis-like structures. Finally, no cytopathic effects, inclusion bodies, and viral DNA replication were detected in infected Bm-5, Se-1, and Cf-124T cell lines. Therefore, these cell lines were accepted as non-productive. This is the first study on the host specifity of ManeNPV in vitro. Our results suggest that Tn5 and LdElta cell lines can provide an important model for studying ManeNPV in vitro system as in the Md203 cell line system.

Anahtar Kelimeler:

Malacosoma neustria NPV, virus replication, host specificity

Comparative Susceptibilities of Six Insect Cell Lines to Infection by Malacosoma neustria Nucleopolyhedrovirus (ManeNPV)

Keywords:

Malacosoma neustria NPV, virus replication, host specificity,

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Van Beek NAM, Hunghes PR. The response time of insect larvae infected with recombinant baculovirus. J Invertebr Pathol 72: 338-347, 1998.
McCutchen BF, Choudary PV, Crenshaw R et al. Development of a recombinant baculovirus expressing an insect-selective neurotoxin: potential for pest control. Biotechnology (NY) 9: 848-852, 1991.
Miller LK, Lu A. The molecular basis of baculovirues host range. In: Miller LK. ed. The Baculoviruses. Plenum Press, New York; 1997: pp. 217-235.
Stewart LM, Hirst M, Lopez Ferber M et al. Construction of an improved baculovirus insecticide containing an insect-specific toxin gene. Nature 352: 85–88, 1991.
Herniou EA, Olszewski JA, Cory JS et al. The genome sequence and evolution of baculoviruses. Ann Rev Entomol 48: 211-234, 2003.
Hostetter DL, Puttler B. A new broad host spectrum nuclear polyhedrosis virus isolated from a celery looper, Anagrapha falcifera (Kirby) (Lepidoptera: Noctuidae). Environ Entomol 20: 1481-1488, 1991. Castro MEB, Souza ML, Arauja S et al. Replication of Anticarsia gemmatalis nuclear polyhedrosis virus four lepidopteran cell lines. J Invertebr Pathol 69: 40-45, 1997. Lynn DE. Comparative susceptibilities of twelve cell lines to infection by three baculoviruses. J Invertebr Pathol 82: 129-131, 2003.
Demir I, Demirbag Z. A productive replication of Hyphantria cunea nucleopolyhedrovirus in Lymantria dispar cell line. J Microbial Biotechnol 16(10): 1485-1490, 2006.
Jankevica J, Cudare Z, Ose V. New isolate of Malacosoma nesutria nuclear polyhedrosis virus in Latvia. J Invertebr Pathol 71: 283-285, 1998.
Kikhno IM, Strokovskaya LI. Physical mapping of Malacosoma neustria nuclear polyhedrosis virus genome. Biopolimery i Kletka 13(3): 218-221, 1997.
Yaman M, Nalcacioglu R, Belduz AO et al. A nuclear polyhedrosis virus of the lackey moth, Malacosoma neustria (Lepidoptera: Lasiocampidae) in Turkey. IOBC wprs Bulletin 26(1): 233-236, 2003.
Mitsuhashi J. A continuously growing cell line from larval hemocytes of Malacosoma neustria testacea. Jpn J Ent 64(3): 692-699, 1996.
Demir I, Gurel N, Nalcacioglu R et al. Productive replication of Malacosoma neustria nucleopolyhedrovirus (ManeNPV) in Md203 cell line. Turk J Biol 33: 239-248, 2009.
Brown M, Faulkner P. Factors affecting the yield virus in a cloned cell line of Trichoplusia ni infected with a nuclear polyhedrosis virus. J Invertebr Pathol 26: 251-257, 1975.
Reed L, Muench H. A simple method for estimating fifty percent end points. Am J Hyg 27: 493-497, 1938.
Invitrogen. MaxBac 2.0 Transfection and expression manual. In: Invitrogen Corporation, A Manual of Methods for Baculovirus Expression. 3985 B Sorrento Valley Blvd, San Diego; USA, 1996: pp. 35-36.
Woo SD. Rapid detection of multiple nucleopolyhedroviruses using polymerase chain reaction. Mol Cells 11: 334-340, 2001.
Keddie AB, Erlandson MA, Hilchie GJ. Establishment and characterization of three Malacosoma disstria cell lines. J Invertebr Pathol 66: 136-142, 1995.
Blissard GW. Baculovirus-insect cell interactions. In: Vlak JM, De Gooijer CD, Tramper J et al. eds. Insect Cell Cultures: Fundamental and Applied Aspects. Kluwer Academic Publishers, P.O. Box 17,3300, AA Dordrecht; The Netherlands, 1996: pp. 73-93.
Williams GV. Faulkner P.
Replication patterns and
cytopathology of cells infected with baculoviruses. In: Vlak JM,
De Gooijer CD, Tramper J et al. eds. Insect Cell Cultures:
Fundamental and Applied Aspects, Kluwer Academic
Publishers, P.O. Box 17,3300, AA Dordrecht; The Netherlands, 1996: pp. 95-110.
Demirbag Z, Belduz AO, Erturk O et al. Comparative replication of Autographa californica nuclear polyhedrosis virus in insect cell cultures. Turk J Biol 21: 63-70, 1997.
Harrison RL, Lynn DE. New cell lines derived from the black cutworm, Agrotis ipsilon, that support replication of the A. ipsilon multiple nucleopolyhedrovirus and several groupI nucleopolyhedroviruses. J Invertebr Pathol (in press), 2008.
Levin DB, Huang J.
Spodoptera littoralis type B
nucleopolyhedrovirus infection of a grasshopper cell line. J
Invertebr Pathol 74: 184-192, 1999.
Shim H J, Roh JY, Choi JY et al. Isolation and characterization of a Lymantria dispar multinucleocapsid nucleopolyhedrovirus isolate in Korea. J Microbiol 41(4): 306-311, 2003.
Shirata N, Ikeda M, Kamiya K et al. Replication of nucleopolyhedrosisviruses of Autographa californica (Lepidoptera: Noctuidae), Bombyx mori (Lepidoptera: Bombycidae), Hyphantria cunea (Lepidoptera: Arctiidae) and Spodoptera exigua lepidopteran cell lines. Appl Entomol Zool 34: 507-516, 1999.
Lynn DE. Improved efficiency in determining the titer of the Autographa californica baculovirus nonoccluded virus. Bio Techniques 13: 282-285, 1992.
McIntosh AH, Ignoffo CM, Andrews PL. In vitro host range of five baculovirus in lepidopteran cell lines. Intervirology 23: 150:156, 1985.
Riegel CI, Slavicek JM. Characterization of the replication cycle of the Lymantria dispar nuclear polyhedrosis virus. Virus Res 51: 9-17, 1997.
McClintock JT, Dougherty EM, Wiener RM. Semipermissive replication of a nuclear polyhedrisis virus of Autographa californica in a gypsy moth cell line. J Virol 57: 197-204, 1986.
Clem RJ, Miller LK. Apoptosis reduces both the in vitro replication and the in vivo infectivity of a baculovirus. J Virol 67: 3730-3738, 1993.
Razvi ES, Welsh, RM. Apoptosis in viral infections. Ads Virus Res 45: 1-60, 1995.
Hang X, Dong W, Guarino LA. The lef-3 gene of Autographa californica nuclear polyhedrovirosis virus encodes a single- stranded DNA-binding protein. J Virol 69: 3924-3928, 1995.
Kool M, van den Berg PM, Tramper J et al. Location of two pupative origins of DNA replication of Autographa californica nuclear polyhedrosis virus. Virology 192: 94-101, 1993.
Grace TDC. Insect cell culture and virus research. In Vitro 3: 104-117, 1967.
Vaughn JL, Goodman RM, Tomkins GT et al. The establishment of two cell lines from the insect Spodoptera frugiperda (Lepidoptera, Noctiudae). In Vitro 13: 213-217, 1977.
Gelernter WD, Federici BA. Continuous cell line from Spodoptera exigua (Lepidoptera: Noctuidae) that supports replication of nuclear polyhedrosis viruses from Spodoptera exigua and Autographa californica. J Invertebr Pathol 48:199– 207, 1986.
Bilimoria SL, Sohi SS. Development of an attached strain from a continuous insect cell line. In Vitro 13: 461-466, 1977.