Although current drug regimens for HIV infection are prolonging millions of lives, CD4+ T lymphocytes harboring latent provirus remain an obstacle to a sterilizing cure. To develop and test clinical interventions, new blood tests are needed that are sensitive enough to detect extremely rare latently-infected cells and accurate enough to measure changes over time. To fully characterize the response to treatment interventions, methods to analyze the properties of single latent cells are also needed. In this project, a micro-fabricated cell culture platform will be used to individually identify and characterize latently-infected cells. Primary lymphocytes will be cultivated in arrays of 105 to over 107 micro-wells, and latent cells will be identified by p24 production after stimulation with anti-CD3/28. After optimization, the sensitivity and reproducibility of this latency assay will be assessed and compared with established assays for HIV DNA and cell-associated infectivity. If the R21 phase succeeds in developing a quantitative assay for replication-competent latent virus, the same platform will be used in the R33 phase to characterize individual latently-infected cells in unprecedented detail, by combining detection based on p24 with measurements of cell-surface phenotype, secreted cytokines, and HIV DNA sequences. Single-cell analyses using these correlated measurements will quantitatively assess the importance of replication-defective proviruses and of homeostatic proliferation.

Public Health Relevance

This research is relevant to public health since it will develop a new test for residual HIV in the 700,000 patients currently receiving effective antiretroviral therapy. Laboratory tests for this remaining virus are essential for the development and testing of treatments to cure HIV. The proposed research is therefore highly relevant to the part of the NIAID's mission that pertains to the better understanding and treatment of infectious diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21AI104283-02
Application #
8631036
Study Section
Special Emphasis Panel (ZAI1-JRR-A (S1))
Program Officer
Stansell, Elizabeth H
Project Start
2013-03-06
Project End
2015-02-28
Budget Start
2014-03-01
Budget End
2015-02-28
Support Year
2
Fiscal Year
2014
Total Cost
$189,575
Indirect Cost
$53,386
Name
Veterans Medical Research Fdn/San Diego
Department
Type
DUNS #
933863508
City
San Diego
State
CA
Country
United States
Zip Code
92161