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Updated: Sep 16, 2025

Single-cell Quantitation of mRNA and Surface Protein Expression in Simian Immunodeficiency Virus-infected CD4+ T Cells Isolated from Rhesus macaques
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High-throughput High Content Quantification of HIV-1 Viral Infectious Output.

Teresa LuPone1, Alexis Brantly1,2, Oluwatofunmi Oteju3

  • 1Department of Microbiology and Immunology, Drexel University College of Medicine, Philadelphia, PA 19102.

Biorxiv : the Preprint Server for Biology
|July 9, 2025
PubMed
Summary
This summary is machine-generated.

A new high-content imaging assay allows for rapid, accurate assessment of human immunodeficiency virus type 1 (HIV-1) infection dynamics. This method correlates infectivity with p24 secretion, improving understanding of viral replication and pathogenesis.

Keywords:
HIV-1high content imaginginfectivityp24 secretionviral titer

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Area of Science:

  • Virology
  • Cell Biology
  • Biotechnology

Background:

  • Human immunodeficiency virus type 1 (HIV-1) infection remains a global health concern, leading to significant pathology and comorbidities.
  • Antiretroviral therapy (ART) suppresses viral replication but is often delayed, allowing unchecked viral progression and development of comorbidities.
  • Current methods for evaluating HIV-1 infection dynamics in vitro lack efficiency for large-scale, direct infectivity assessments.

Purpose of the Study:

  • To develop and validate a high-content imaging reporter assay for rapid, reproducible evaluation of HIV-1 infection kinetics.
  • To enable direct correlation between viral infectivity and other measures like p24 secretion.
  • To provide a more robust understanding of HIV-1 replication dynamics in distinct cell populations.

Main Methods:

  • Utilized modified human osteosarcoma cells (GHOST cells) engineered to fluoresce upon HIV-1 infection.
  • Implemented a high-content imaging and analysis pipeline for rapid, large-scale sample evaluation.
  • Integrated the assay with existing experimental workflows analyzing p24 secretion in culture supernatants.

Main Results:

  • The developed assay provides rapid, consistent, and replicable evaluation of HIV-1 infectivity.
  • Demonstrated a direct correlation between fluorescence-based infectivity measurements and p24 secretion levels.
  • The assay integrates seamlessly with established methods for analyzing viral markers.

Conclusions:

  • The high-content imaging reporter assay offers a high-throughput method for assessing HIV-1 direct infectivity.
  • This assay enhances the interrogation of HIV-1 infection kinetics and facilitates deeper understanding of viral pathogenesis.
  • The method supports the evaluation of infectivity changes across various treatments and timepoints.