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Updated: Mar 30, 2026
A High-throughput Cre-Lox Activated Viral Membrane Fusion Assay to Identify Inhibitors of HIV-1 Viral Membrane Fusion
Published on: August 14, 2018
Anti-HIV-1 activity determined by β-galactosidase activity in the multinuclear activation of an indicator assay is
Fusako Miyamoto1, Kumi Kawaji1, Shinya Oishi2
1Division of Miyagi Community Health Promotion, Tohoku University Graduate School of Medicine and Tohoku Medical Megabank Organization, Sendai, Japan Division of Infection Control and Laboratory Diagnostics, Department of Internal Medicine, Tohoku University Graduate School of Medicine, Sendai, Japan.
Background:
Direct comparison of enzymatic and original blue cell-counting detections with the multinuclear activation of an indicator (MAGI) cells, so far, remains to be performed in parallel. Although inhibitors for reverse transcription solely inhibit the reverse transcription step, those for HIV-1 entry block syncytium formation of HIV-1-infected MAGI cells in addition to the entry (dual inhibition). It raises a concern that reduction of enzymatic activity is artificially influenced by syncytium-blocking activity of inhibitors for entry.
Methods:
The MAGI cells with a syncytium inducible strain, HIV-1IIIB, were used for anti-HIV activity determination both with conventional counting with X-Gal staining and measurement of chlorophenol red β-d-galactopyranoside conversion with a plate reader.
Results:
Infectivity of HIV-1 in the MAGI cells was highly correlated with both methods. In microscopic observation, small blue cells with single or a couple of nuclei were dominantly observed in the presence of inhibitors for entry, but not in the presence of those for reverse transcription. Actual anti-HIV-1 activities were comparable or moderately sensitive in the chlorophenol red β-d-galactopyranoside method.
Conclusions:
Antiviral activities of inhibitors for entry obtained from both enzymatic and counting methods appear to be comparable, even in infection of a highly syncytia inducible HIV-1IIIB strain.
