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Human immunodeficiency virus type 1 spinoculation enhances infection through virus binding.
U O'Doherty1, W J Swiggard, M H Malim
1Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104-6148, USA.
Journal of Virology
|October 12, 2000
Summary
Centrifugal inoculation, or spinoculation, significantly enhances human immunodeficiency virus type 1 (HIV-1) infection efficiency in vitro by increasing viral particle binding to target cells. This method accelerates HIV-1 research and improves virus-based therapies.
Area of Science:
- Virology
- Cell Biology
- Molecular Biology
Background:
- Studying early human immunodeficiency virus type 1 (HIV-1) events in vitro is challenging due to low synchronous infection rates.
- Centrifugal inoculation (spinoculation) improves HIV-1 infection efficiency, but its mechanism remains unclear.
Purpose of the Study:
- To elucidate the mechanism by which spinoculation enhances HIV-1 infection in vitro.
- To quantify viral particle binding, association with cells, and reverse transcript formation during spinoculation.
Main Methods:
- Real-time PCR was used to quantify viral particles.
- HIV-1(IIIB) virions were centrifuged onto CEM-SS T-cell targets at 1,200 x g for 2 hours at 25°C.
- Infection efficiency was assessed by p24(gag) expression after 24 hours of culture.
Main Results:
- Spinoculation increased HIV-1 particle binding to T-cells by approximately 40-fold compared to simple mixing.
- Reverse transcript levels per target cell were similarly enhanced after spinoculation.
- Approximately 100% of target cells showed productive infection after 24 hours of culture.
Conclusions:
- Spinoculation enhances HIV-1 infection by depositing virions onto target cells, overcoming diffusion limitations in viral adsorption.
- This technique facilitates the study of post-entry HIV-1 infection steps.
- Spinoculation holds promise for enhancing the efficacy of virus-based genetic therapies.