Methods for enhancing the production of viral vaccines in PKR-deficient cell culture

Abstract

Methods for enhancing the production of viral vaccines in animal cell culture are described. These methods rely on the manipulation of the cellular levels of certain interferon induced antiviral activities, in particular, cellular levels of double-stranded RNA (dsRNA) dependent kinase (PKR), PKR-deficient cells are obtained by a) transfecting a parent cell line with a PKR antisense polynucleotide; b) unaided uptake into a cell line by culturing said cell line in the presence of a PKR antisense polynucleotide; c) transfection of a parent cell line with a PKR dominant negative mutant; or d) culturing a cell line in the presence of 2-aminopurine. In cell cultures deficient for PKR, viral yield is enhanced by several orders of magnitude over cell cultures with normal levels of these proteins making these cell cultures useful for the production of viral vaccines.