Transformed lepidopteran cells expressing a protein of the silkmoth fat body display enhanced susceptibility to baculovirus infection and produce high titers of budded virus in serum-free media.
Baculovirus vectors constitute important tools for therapeutic protein production and mammalian cell transduction for gene therapy applications. A prerequisite for such applications is that the cell lines in which baculoviruses are propagated be maintained in serum-free media that are devoid of potential human pathogens. However, in serum-free media, the performance of baculovirus-based systems can be significantly reduced. In this report, we show that silkmoth-derived host cell lines for the Bombyx mori-nuclear polyhedrosis virus (BmNPV) that are transformed with the gene for the promoting protein (PP), a silkmoth-derived secreted factor containing a lipid-binding domain, display enhanced susceptibility to BmNPV infection and enhanced budded virus productivity in serum-free media. For transformed silkmoth cells maintained in serum-free media, the rate of BmNPV entry is enhanced by two orders of magnitude relative to the untransformed cells, while the rate of budded virus production is increased five-fold. The infectivity-enhancing effect can be also conferred to normal cells grown in serum-free media by addition of conditioned media from the transformed cells, which contain the secreted recombinant PP. Thus, PP substitutes for serum factors whose presence facilitates baculovirus entry into the cells. However, the effects of silkmoth-derived PP may be specific to the BmNPV-silkmoth system since little or no changes in viral infectivity are obtained by PP expression in Trichoplusia ni-derived High-Fivetrade mark cells grown in serum-free media and infected with a different baculovirus (AcNPV).
Iatrou
Kostas
K
Institute of Biology, Demokritos National Centre for Scientific Research, P.O. Box 60228, Patriarchou Grigoriou & Neapoleos Str., 153 10 Aghia Paraskevi Attikis, Athens, Greece. iatrou@bio.demokritos.gr
Swevers
Luc
L
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D012831:Bombyx / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D018929:Cell Culture Techniques; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral; D017077:Culture Media, Conditioned / Q000737:chemistry / Q000494:pharmacology; D016895:Culture Media, Serum-Free / Q000737:chemistry / Q000494:pharmacology*; D004198:Disease Susceptibility / Q000821:virology; D005216:Fat Body / Q000378:metabolism; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017924:Nucleopolyhedroviruses / Q000502:physiology*; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects*
D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D019476:Insect Proteins; D011994:Recombinant Proteins