HIV latency is emerging as barrier to eradicating HIV-1 infection. The current theoretical paradigm for eliminating the viral reservoir is known as `Shock and Kill', reactivation of latent virus using non-targeted small molecules. However, significant hurdles will have to be overcome for this approach to be successful. These include stochastic viral reactivation, avoidance of global T-cell activation, and limited cellular death upn re- activation. In addition to these limitations, most of what we know about latency is derived from the study of latency in peripheral blood mononuclear cells. Using a newly developed dual fluorescence reporter HIV, we have observed that latency was significantly more prevalent in CD4 T cells isolated from oral lymphoid tissues (tonsil) than in CD4 T cells from peripheral blood or spleen. In addition, latency in CD4 T cells from oral lymphoid tissues was strongly dependent on the presence of non-CD4 T cells. Thus, the biology of latency and its reactivation may differ in primary oral lymphoid tissues when compared to other lymphoid tissues. If so, therapeutic interventions might need to be modified to account for the unique characteristics of latency in oral lymphoid tissues. We propose to use a multi-pronged, systems-biology strategy to further define HIV latency and its mechanism in primary oral lymphoid tissues. We will use our new HIV-1 reporter to identify, quantify, and purify latently infected cells in their native state earl after infection of tonsil CD4 T cells. Purified latently infected cells will be characterized usingthe following tools: 1. CyTOF single-cell analysis of cell-surface markers, T-cell signaling networks, and T-cell effector function; 2. RNAseq and microRNAseq analysis of the cellular transcriptome; 3. Mechanistic studies using RNP-Cas9 gene interference to identify key regulators of HIV latency in primary oral lymphoid tissues. In addition, we will define the relative contribution of intercellular interactions and secreted factors in the ability of non-CD4 T cells to influence HIV latency in CD4 T cells in oral lymphoid tissues. By combining our dual-fluorescence latency model with powerful systems biology techniques and the tonsil HLAC system, we expect to significantly increase our understanding of HIV latency and to identify potential new therapeutic opportunities.

Public Health Relevance

HIV-1 persists in spite of antiretroviral therapy due to a reservoir of long-lived and latently infected resting memory CD4+ T-cells. Using a new HIV reporter tool to study latency, we have recently observed that latency was significantly more prevalent in CD4 T cells isolated from oral lymphoid tissues (tonsil) than in CD4 T cells from peripheral blood or spleen. In addition, latency in CD4 T cells from oral lymphoid tissues was strongly dependent on the presence of non-CD4 T cells. Thus, the biology of latency and its reactivation may differ in primary oral lymphoid tissues when compared to other lymphoid tissues. If so, therapeutic interventions might need to be modified to account for the unique characteristics of latency in peripheral lymphoid tissues, including primary oral lymphoid tissues. We will use a multi-pronged, systems-biology strategy to further define HIV latency and its mechanism in primary oral lymphoid tissues. We will use our new HIV-1 reporter to identify, quantify, and purify latently infected cells in their native state early after infection. By combinng our dual-fluorescence latency model with powerful systems biology techniques and the tonsil HLAC system, we expect to significantly increase our understanding of HIV latency and to identify potential new therapeutic opportunities.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
5R01DE026010-05
Application #
9624757
Study Section
Special Emphasis Panel (ZDE1)
Program Officer
Gannot, Gallya
Project Start
2016-04-01
Project End
2021-01-31
Budget Start
2019-02-01
Budget End
2020-01-31
Support Year
5
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Buck Institute for Age Research
Department
Type
DUNS #
786502351
City
Novato
State
CA
Country
United States
Zip Code
94945
Battivelli, Emilie; Dahabieh, Matthew S; Abdel-Mohsen, Mohamed et al. (2018) Distinct chromatin functional states correlate with HIV latency reactivation in infected primary CD4+ T cells. Elife 7:
Besnard, Emilie; Hakre, Shweta; Kampmann, Martin et al. (2016) The mTOR Complex Controls HIV Latency. Cell Host Microbe 20:785-797