High-throughput high content quantification of HIV-1 viral infectious output

Area of Science:

• Virology
• Cell Biology
• Immunology

Background:

• Human immunodeficiency virus (HIV-1) infection is a global health concern, leading to significant pathology and comorbidities.
• Antiretroviral therapy (ART) suppresses viral replication but doesn't prevent HIV-1-associated comorbidities, indicating gaps in understanding disease pathogenesis.
• Current methods for studying HIV-1 replication are insufficient for rapid, high-throughput analysis, hindering the development of targeted therapies.

Purpose of the Study:

• To develop and validate a novel high-content imaging reporter assay for assessing HIV-1 replication dynamics.
• To enable rapid, consistent, and replicable evaluation of HIV-1 infectivity and kinetics.
• To correlate viral infectivity with p24 secretion for a deeper understanding of HIV-1 pathogenesis.

Main Methods:

• Utilized modified human osteosarcoma cells (GHOST cells) engineered to fluoresce upon HIV-1 infection.
• Integrated a high-content imaging approach for rapid sample evaluation.
• Combined fluorescence detection with p24 secretion analysis from cell supernatants.

Main Results:

• The developed assay provides rapid and reproducible assessment of HIV-1 infection.
• The assay allows for direct correlation between HIV-1 infectivity and p24 antigen secretion.
• This method enhances the interrogation of HIV-1 replication kinetics in distinct cell populations.

Conclusions:

• The novel reporter assay offers a robust platform for studying HIV-1 replication dynamics.
• This tool facilitates a more comprehensive understanding of the relationship between viral replication and HIV-1-associated pathogenesis.
• The assay can be readily integrated into existing experimental workflows for improved HIV-1 research.