4th International AIDS Conference


Stockholm, Sweden. — June 12-16, 1988

[TITLE:] UTILITY OF A HIV 1 RETROVIRAL VECTOR SYSTEM FOR GENE TRANSFER INTO HUMAN CELLS

Int Conf AIDS. 1988 Jun 12-16;4:1.113 (abstract no. 1002)

V. Heisig, G. Jahn, M. Ebeling, R. Laufs
Inst. f. Med. Mikrobiologie and Immunologie Universität Hamburg, Hamburg, W. Germany

Retroviral vectors have been constructed for gene transfer in mammalian and avian cells. However, they are restricted in host range. We describe a retroviral vector system based entirely on an human immunodeficiency virus (HIV 1) containing the ability to carry out efficient gene transfer into human cells.

By establishing a helper cell line that produces the trans-acting viral gene products we propagate the cis-acting components in them and harvest defective viral particles that contain only the cis-acting components. In HIV 1 the packaging signal has not been identified. We postulate that the sequence for the packaging signal is located between the primer binding site (PBS) and the gag genes. Therefore we constructed deletion mutants in this region and transfected this mutants into H9 cells. The plasmid pVH3H containing the complete proviral genome of HIV 1 and a hygromycin B gene for selection was used for constructions. An additional deletion in the 3 'LTR (-138 to -48) has been introduced into all plasmids. We have established cell lines with the mutants and studied the reverse transcriptase activity, RNA and protein synthesis.

The cis-acting component, a retroviral self inactivating vector containing two HIV 1 LTR's was transfected into the packaging defective helper cell lines. This plasmid contains the hygromycin B gene between both LTR's. Preliminary analysis of the supernatants of some cell lines indicate that defective particles are produced and transferred to human cell lines.

Such a human retrovirus gene transfer system could be useful for the transfer of HIV 1 genes into human cells.

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