Description of Research Expertise:
Key words: HIV, latency, reservoirs, dendritic cells, viral pathogenesis, proviral integration, retrovirus, virology, T cell activation, resting T cells.
Description of Research
Highly active anti-retroviral therapy can clear the blood of HIV-1 virions. But, despite long-term suppression of virus, when the drugs are stopped the virus returns. Thus, reservoirs of latent, treatment-resistant HIV-1 exist in infected individuals and are a major barrier to cure. With the advent of ART, the challenge in the field of HIV is to clear the remaining reservoir. The challenge is significant since the reservoir is very small - less than 1 in a million CD4+ T cells are true HIV reservoir cells. Moreover, identifying the true reservoir is made more difficult because it is hidden among many defective HIV proviruses.
To better understand the challenge to cure HIV, we quantify the reservoir and to measure how much reservoir expression occurs at baseline and to what extent the reservoir visibility can be increased by stimulation. A major hurdle that we face in these studies is to distinguish replication competent HIV from defective proviruses. We believe that we have identified a technology that largely overcomes this hurdle. Our approach utilizes Fiber-optic Array Scanning Technology (or FAST) to identify rare cells that can express HIV proteins at high levels. The technique is largely based on approaches for rare cancer cell detection.
Our lab also studies HIV reservoirs by using an in vitro model that we developed and characterized. We have shown that this model mimics many important aspects of HIV in vivo. This model allows us probe the important differences between the latent and productive state of HIV infection. We are keenly interested to probe why HIV expression is less efficient in latently infected cells. This is an underexplored area that can be addresses with current sequencing and proteomic approaches.
1. Characterize the ability of FAST technology to distinguish and quantify replication competent proviruses from defective proviruses.
2. Characterize the frequency of replication competent proviruses in different patient populations.
3. Probe the mechanistic differences between latent and productive HIV Infection.
Jake VanBelzen - Postbaccalaureate student
Marilia Pinzone- Postdoctoral fellow
Villa C, Porturas T, Sell M, Hwang W, Wall M, DeLeo G, Fetters J, Mignono S, Irwin L: Rapid prediction of stem cell mobilization using volume and conductivity data from automated hematology analyzers. Transfusion : 2018.
Tapia G, Højen J F, Ökvist M, Olesen R, Leth S, Nissen S K, VanBelzen D J, O'Doherty U, Mørk A, Krogsgaard K, Søgaard O S, Østergaard L, Tolstrup M, Pantaleo G: Sequential Vacc-4x and romidepsin during combination antiretroviral therapy (cART): Immune responses to Vacc-4x regions on p24 and changes in HIV reservoirs. The Journal of infection : 2017.
Pinzone Marilia Rita, Graf Erin, Lynch Lindsay, McLaughlin Brigit, Hecht Frederick M, Connors Mark, Migueles Stephen A, Hwang Wei-Ting, Nunnari Giuseppe: Monitoring Integration over Time Supports a Role for Cytotoxic T Lymphocytes and Ongoing Replication as Determinants of Reservoir Size. Journal of virology 90 (23): 10436-10445,2016.
Mavigner Maud, Lee S Thera, Habib Jakob, Robinson Cameron, Silvestri Guido, O'Doherty Una: Quantifying integrated SIV-DNA by repetitive-sampling Alu-gag PCR. Journal of virus eradication 2 (4): 219-226,2016.
Lee Sulggi A, Bacchetti Peter, Chomont Nicolas, Fromentin Remi, Lewin Sharon R, O'Doherty Una, Palmer Sarah, Richman Douglas D, Siliciano Janet D, Yukl Steven A, Deeks Steven G: Anti-HIV Antibody Responses and the HIV Reservoir Size during Antiretroviral Therapy. PloS one 11 (8): e0160192,2016.
Imamichi Hiromi, Dewar Robin L, Adelsberger Joseph W, Rehm Catherine A, O'Doherty Una, Paxinos Ellen E, Fauci Anthony S: Defective HIV-1 proviruses produce novel protein-coding RNA species in HIV-infected patients on combination antiretroviral therapy. Proceedings of the National Academy of Sciences of the United States of America 113 (31): 8783-8,2016.
DeMaster Laura K, Liu Xiaohe, VanBelzen D Jake, Trinité Benjamin, Zheng Lingjie, Agosto Luis M, Migueles Stephen A, Connors Mark, Sambucetti Lidia, Levy David N, Pasternak Alexander O: A subset of CD4/CD8 double negative T cells expresses HIV proteins in patients on ART. Journal of virology 90 (5): 2165-79,2015.
Pasternak Alexander O, DeMaster Laura K, Kootstra Neeltje A, Reiss Peter, O'Doherty Una: Minor Contribution of Host-HIV Readthrough Transcripts to the Level of HIV Cell-associated gag RNA. Journal of virology 90 (2): 1148-51,2015.
Procopio Francesco Andrea, Fromentin Rémi, Kulpa Deanna A, Brehm Jessica H, Bebin Anne-Gaelle, Strain Matthew C, Richman Douglas D, O'Doherty Una, Palmer Sarah, Hecht Frederick M, Hoh Rebecca, Barnard Richard J O, Miller Michael D, Hazuda Daria J, Deeks Steven G, Sékaly Rafick-Pierre: A Novel Assay to Measure the Magnitude of the Inducible Viral Reservoir in HIV-infected Individuals. EBioMedicine 2 (8): 872-81,2015.
Cockerham Leslie R, Siliciano Janet D, Sinclair Elizabeth, O'Doherty Una, Palmer Sarah, Yukl Steven A, Strain Matt C, Chomont Nicolas, Hecht Frederick M, Siliciano Robert F, Richman Douglas D: CD4+ and CD8+ T Cell Activation Are Associated with HIV DNA in Resting CD4+ T Cells. PloS one 9 (10): e110731,2014.
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