Journal of General Virology, Vol 79, 231-237, Copyright © 1998 by Society for General Microbiology
Development of cell lines stably expressing human immunodeficiency virus type 1 proteins for studies in encapsidation and gene transfer
D Haselhorst, JF Kaye and AM Lever
University of Cambridge Department of Medicine, Addenbrooke's Hospital, UK.
Experiments were done to test cell lines for their capacity to express
human immunodeficiency virus type 1 (HIV-1) proteins in a stable manner.
Marked differences were seen in the ability to stably express and export
viral Gag and Pol proteins. Two cell lines, one suspension (MDS) and one
monolayer (SW480), were established which exported these proteins at high
level. Two other cell lines, HeLa and THP-1, showed poorer expression and
very limited particle release. Single cell cloning was used to select the
optimal producing clones from the lines. These produced large quantities of
viral core particles pelletable from the supernatants. Cell lines were
constructed from these clones which stably expressed in addition either the
HIV-1 Envelope or a packageable HIV-based vector. The vector was shown to
be packaged within the viral core particles. Transient transfection of
envelope expressing constructs into a gag-pol plus vector cell line, or the
vector into a gag-pol plus envelope expressing cell line resulted in gene
transfer to CD4+ target cells. These cell lines provide useful tools with
which to study the assembly and export of viral proteins and RNA, for assay
of alternative envelope proteins to pseudotype HIV cores, for assessment of
antiviral drugs and as a source of correctly processed proteins for
immunological studies.
