Abstract
Three insect cell lines were tested for susceptibility to baculovirus infection by use of a typical endpoint assay procedure. Cell lines from Spodoptera frugiperda (IPLB-Sf21AE), Lymantria dispar (IPLB-LdEIta), and Heliothis virescens (IPLB-HvE6s) in 96-well tissue culture plates were each infected with dilutions of extra cellular virus suspensions of the Autographa californica nucleopolyhedrovirus (AcMNPV). In addition, the L. dispar and H. virescens cells were also infected with L. dispar nucleopolyhedrovirus, and Helicoverpa zea nucleopolyhedrovirus, respectively. Each cell/virus combination was incubated at three temperatures: 22, 27 and 32 °C and wells were scored for positive infection (presence of occlusion bodies in cell nuclei) at 2 to 4 day intervals for up to 4 weeks. The resulting data were analyzed by the Spearman-Kärber method, providing virus titers for each combination of virus, cell line, and temperature. The results were categorized by accuracy (assuming the highest titer achieved was the most accurate) and by rapidity of maximum titer. AcMNPV reached the highest titer in each line at 22 °C although equivalent titers were reached with both AcMNPV and HzSNPV in the HvE6a line at all three temperatures. This line actually reported about 100-fold less AcMNPV than the other two lines with the same virus sample. Alternatively, the Sf21AE and LdEIta lines reached 10-fold higher titers at the lowest temperature as compared with the higher temperatures, although also at a slower rate.
Similar content being viewed by others
References
Hughes PR, Wood HA (1986). In vivo and in vitro bioassay methods for baculoviruses. In: Granados RR, Federici BA (eds.), Biology of the Baculovirus, Vol. II, Practical Application for Insect Control. Boca Raton, FL: CRC Press, pp. 1-30.
Lynn DE (1992). A BASIC computer program for analyzing endpoint assays. BioTechniques 12: 880-881.
Lynn DE (1992). Improved efficiency in determining the titer of the Autographa californica baculovirus nonoccluded virus. BioTechniques 13: 282-285.
Lynn DE (1997). Infectivity of different baculovirus phenotypes in insect cell cultures. In Vitro Cell Dev Biol - Animal 33: 19A.
Lynn DE (1999). Comparison of cell line maintenance procedures on insect cells used for producing baculoviruses. In Vitro Cell Dev Biol - Animal 35: 248-251.
Lynn DE (1999). Development of insect cell lines: virus susceptibility and applicability to prawn cell culture. Methods in Cell Science 21: 173-181.
Lynn DE, Dougherty EM, McClintock JT, Loeb M (1988). Development of cell lines from various tissues of Lepidoptera. In: Kuroda Y, Kurstak E, Maramorosch K (eds.), Invertebrate and Fish Tissue Culture. Tokyo: Japan Scientific Societies Press, pp. 239-242.
Lynn DE, Dougherty EM, McClintock JT, Shapiro M (1989). Comparative replication of Lymantria dispar nuclear polyhedrosis virus strains in three continuousculture cell lines. Appl Environ Microbiol 55: 1049-1051.
Lynn DE, Shapiro M (1998). New cell lines from Heliothis virescens: characterization and susceptibility to baculoviruses. J Invertebr Pathol 72: 276-280.
Vaughn JL, Goodwin RH, Tompkins GJ, McCawley P (1977). The establishment of two cell lines from the insect Spodoptera frugiperda (Lepidoptera: Noctuidae). In Vitro 13: 213-217.
Volkman LE, Goldsmith PA (1982). Generalized immunoassay for Autographa californica nuclear polyhedrosis virus infectivity in vitro. Appl Environ Microbiol 44: 227-233.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Lynn, D.E. Effects of temperature on the susceptibility of insect cells to infection by baculoviruses. Methods Cell Sci 23, 221–225 (2001). https://doi.org/10.1023/A:1016394421408
Issue Date:
DOI: https://doi.org/10.1023/A:1016394421408