Enhancer-deficient amphotropic murine leukemia virus and recombinants with heterologous transcription elements can be efficiently amplified and detected in Mus dunni fibroblasts

Abstract

Amphotropic murine leukemia virus (MLV) replicates in cells from various mammalian species, including humans, and is a potential contaminant in MLV vector preparations for human gene transfer studies. Mus dunni fibroblasts are routinely used for amplification and detection of contaminating virus. We have recently characterized an amphotropic MLV mutant lacking the 75-bp viral enhancer elements and spontaneous MLV-(RCMV) recombinants that have acquired cytomegalovirus (CMV) transcription elements. Both of these viruses replicate in specific human cell types. To test whether the formation of such viruses can be detected and controlled with current routine procedures, we have analyzed the replication of these amphotropic MLV mutants in Mus dunni fibroblasts. We find that M. dunni cells are permissive for enhancer-deficient and CMV promoter-recombinant MLV from several human cell lines. Thus, M. dunni fibroblasts are suitable for the amplification and subsequent detection of enhancer-deficient and enhancer-recombinant MLV in vector preparations.