Baculovirus interaction with nontarget organisms: a virus-borne reporter gene is not expressed in two mammalian cell lines

Abstract

The safety of baculoviruses with respect to mammalian species was studied by using a genetically engineered recombinant of Autographa californica nuclear polyhedrosis virus. This recombinant contains the chloramphenicol acetyltransferase (CAT) gene under the control of a mammalian-active promoter and expresses substantial levels of CAT activity on infection of permissive and nonpermissive insect cells (L.F. Carbonell, M.J. Klowden, and L.K. Miller, J. Virol. 56:153-160, 1985). Extremely low levels of CAT activity were detected in mouse and human cell lines that were continuously exposed to the A. californica nuclear polyhedrosis virus recombinant. The appearance of CAT was not inhibited by cycloheximide. Isopycnic centrifugation of purified inoculum showed that a low level of CAT activity was associated with the insect-derived viral particles. Thus, the observed CAT activity is carried into the cells with the virus inoculum, and active expression of the baculovirus-borne CAT gene is not observed in either cell line. The inability of the CAT gene to be expressed in these cell lines with this model system provides additional assurance of the safety of insect baculoviruses with respect to mammalian species.