Wardrop E, Lamarre D; International Conference on AIDS.
Int Conf AIDS. 1993 Jun 6-11; 9: 148 (abstract no. PO-A05-0084).
Dept. of Biochemistry, Bio-Mega/Boehringer Ingelheim Research Inc., Canada.
A bioassay for the replication of HIV-1 that requires few manipulations has been developed for the rapid quantitation of drug inhibition of acute infection by laboratory strains. The B lymphoblastoid AA-2 cell line expresses a high level of CD4 and is highly permissive for HIV-1 infection. The AA-2 cell line has been stably transfected with a beta-galactosidase gene (lac Z) under the control of a truncated HIV-1 Long Terminal Repeat (LTR-153 to +80) using DNA-mediated gene transfer. Clones displaying an increase in enzyme activity upon infection (15 fold) were selected to set up the bioassay. Assays can be performed in 96-well microtiter plates by infecting a small number of cells (10(4)) with HTLV-IIIB. After a four day incubation beta-galactosidase is readily detectable by the addition of the chromogenic substrate chlorophenol red-beta-D-galactopyranoside in SDS buffer directly to cells in microtiter plates. Reporter activity is sensitive to specific reverse transcriptase and viral protease inhibitors and EC50 curves can be generated by non-linear regression analysis. EC50 determinations obtained with this assay for different protease inhibitors are comparable to values obtained with other assays based on inhibition of syncytia formation and extracellular p24. This assay permits the high capacity screening of antiviral agents directed against any stage of the HIV growth cycle, to study the effect of multi-drug combinations and to characterize the cross-reactivity of antiviral agents against resistant strains.
Publication Types:
Keywords:
- Acquired Immunodeficiency Syndrome
- Antigens, CD4
- Biological Assay
- Cell Line
- HIV Infections
- HIV Long Terminal Repeat
- HIV Seropositivity
- HIV-1
- Lymphocytes
- beta-Galactosidase
- genetics
- immunology
Other ID:
UI: 102202885
From Meeting Abstracts