HIV-1 persistence in viral reservoirs has been the major obstacle for the eradication of HIV-1. Latently infected T cells are one of the potential sources of the virus that is found as residual low-level viremia (LLV) and latent infection of resting memory CD4+ T cells is well demonstrated. Assays have been developed to quantify cells that harbor replication-competent HIV-1 among resting CD4+ T cells in patients on HAART. A viral outgrowth assay is the current gold-standard method to measure latently HIV-1 infected resting CD4+ T cells. However, this assay requires large numbers of resting CD4+ T cells purified from blood and is costly and labor- intensive involving a serial dilution to measure infectious units per million (IUPM) resting CD4+ T cells. To overcome the issues with the current assay, in this application we propose an assay based on next- generation sequencing (NGS) technology. In the proposed assay, the number of different sequences of the viruses bulk-cultured from the resting CD4+ T cells will be directly analyzed to score the number of latently HIV-1-infected CD4+ T cells. Since deep sequencing is highly sensitive to detect multiple variants in a viral population, it will be possible to count the number of distinct viruses that hae been induced to replicate in a bulk culture. These studies will provide valuable information for curing HIV infection and lead to a development of an assay to quantify the latent HIV-1 reservoir with improved accuracy, less time and resource consuming, and less labor intensive.

Public Health Relevance

HIV-1 persistence in viral reservoirs has been the major obstacle for the eradication of HIV-1. Accurate measurement of the size of the HIV-1 latent reservoir is essential for evaluating strategies to eliminate them. Latent HIV-1 infection of restig memory CD4+ T cells is well demonstrated. Development of a simple, rapid, cost-effective, and sensitive assay that can quantify resting CD4+ T cells that harbor replication-competent HIV-1 in patients on highly active antiretroviral therapy will represent important tools in developing cures for HIV infection.

Agency
National Institute of Health (NIH)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21AI113124-01
Application #
8768898
Study Section
Special Emphasis Panel (ZAI1)
Program Officer
Stansell, Elizabeth H
Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599