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Related Experiment Video

Updated: Sep 26, 2025

Phenotypic and Functional Analysis of Activated Regulatory T Cells Isolated from Chronic Lymphocytic Choriomeningitis Virus-infected Mice

Phenotypic and Functional Analysis of Activated Regulatory T Cells Isolated from Chronic Lymphocytic Choriomeningitis Virus-infected Mice

Published on: June 22, 2016

CD8 T Cell Virus Inhibition Assay Protocol.

Yinyan Xu¹, Ann Marie Weideman^2,3, Maria Abad-Fernandez¹

¹Department of Microbiology & Immunology, UNC Chapel Hill School of Medicine, Chapel Hill, North Carolina, USA.

|April 18, 2022

Summary

The viral inhibition assay (VIA) was improved with independent replicate cultures for better reproducibility in measuring CD8+ T cell HIV control. This enhances clinical testing of HIV vaccines and therapies.

Keywords:

CD4 CD8 HIV JRCSF T-cell Virus inhibition p24

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In Vitro Assay to Evaluate the Impact of Immunoregulatory Pathways on HIV-specific CD4 T Cell Effector Function

In Vitro Assay to Evaluate the Impact of Immunoregulatory Pathways on HIV-specific CD4 T Cell Effector Function

Published on: October 15, 2013

Area of Science:

Immunology
Virology
Clinical Research

Background:

The human immunodeficiency virus (HIV)-1 viral inhibition assay (VIA) assesses CD8+ T cell-mediated inhibition of HIV replication in CD4+ T cells.
This assay is crucial for clinical testing of HIV vaccines and immunotherapies, but variations exist in protocols and readouts.
Existing VIA protocols differ in culture duration, CD4+ T cell purity, HIV strains used, and methods for quantifying viral inhibition.

Purpose of the Study:

To describe modifications to a 7-day VIA protocol to improve assay reproducibility.
To introduce independent replicate cultures for HIV-infected CD4+ T cells (HIV-CD4) and HIV-CD4:CD8 T cell cultures.
To identify methodological and analysis changes for enhanced assay performance.

Main Methods:

Modified a 7-day VIA protocol, emphasizing independent replicate cultures for HIV-CD4 and HIV-CD4:CD8 T cell cultures.
Quantified virus inhibition using a ratio of weighted averages of p24+ cells in replicate cultures.
Calculated 95% confidence intervals for inhibition measurements.

Main Results:

Introduction of independent replicate cultures significantly improved assay reproducibility.
Virus inhibition by CD8 T cells in people living with HIV (PLWH) on antiretroviral therapy (ART) remained largely stable over time.
Identified specific methodological and analysis changes applicable to other VIA protocols.

Conclusions:

The modified VIA protocol enhances reproducibility for assessing CD8+ T cell-mediated HIV inhibition.
The stability of CD8 T cell inhibition in PLWH on ART supports the assay's utility for evaluating therapeutic interventions.
These improvements facilitate more reliable clinical testing of HIV vaccines and immunotherapies.