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)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21AI113124-01
Application #
8768898
Study Section
Special Emphasis Panel (ZAI1)
Program Officer
Stansell, Elizabeth H
Project Start
2014-07-01
Project End
2016-06-30
Budget Start
2014-07-01
Budget End
2015-06-30
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