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Bovine Leukaemia Virus Packaging Cell Line for Retrovirus-mediated Gene Transfer

Josef Ban^1,

¹ McArdle Laboratory for Cancer Research
and² Department of Meat and Animal Science, University of Wisconsin, Madison, Wisconsin 53706, U.S.A.

Retroviral packaging cell lines were constructed by using the gag-pol gene of spleen necrosis virus, the gag-pol gene of Moloney murine leukaemia virus and the env gene of bovine leukaemia virus. The plasmids containing the gag-pol genes and the plasmid containing the env gene were cotransfected into NIH/3T3 and D17 cells. The cells containing the helper virus constructs were tested for their ability to package replication-defective murine leukaemia and avian reticuloendotheliosis retrovirus vectors. The titre of vector virus produced by each of the retroviral packaging cell lines was about 10² colony-forming units per ml of medium. Tests for events that might result in intact replication-competent retroviruses showed no evidence for the generation of such viruses. The vector viruses were able to infect dog and rat cells. Bovine cells were infected only after their cocultivation with the retroviral packaging cell lines producing murine leukaemia virus vectors, perhaps as a result of a low concentration of receptors.

Keywords: BLV, MLV, SNV, retrovirus packaging cell line

Present address: Cancer Research Institute of Slovak Academy of Sciences, Bratislava, Czechoslovakia.

Received 13 December 1988; accepted 24 April 1989.