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.
Marilia Pinzone - Postdoctoral fellow
Maria Paola Bertuccio - Postdoctoral fellow
LaMont Cannon - Postdoctoral fellow
Emmanuele Venanzi-Rullo - Infectious Diesease Fellow
Villa CH, Porturas T, Sell M, Wall M, DeLeo G, Fetters J, Mignono S, Irwin L, Hwang WT, O'Doherty U: Rapid prediction of stem cell mobilization using volume and conductivity data from automated hematology analyzers. Transfusion 58 (2): 330-338,2018.
Natesampillai S, Cummins NW, Nie Z, Sampath R, Baker JV, Henry K, Pinzone M, O'Doherty U, Polley EC, Bren GD, Katzmann DJ, Badley AD: HIV Protease-generated Casp8p41, when bound and inactivated by Bcl2, Is degraded by the proteasome. Journal of Virology 92 (13): 2018.
Strongin Z, Sharaf R, VanBelzen DJ, Jacobson JM, Connick E, Volberding P, Skiest DJ, Gandhi RT, Kuritzkes DR, O'Doherty U, Li JZ: Effect of short-term antiretroviral therapy interruption on levels of integrated HIV DNA. Journal of Virology 92 (12): 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, Sommerfelt M A: 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 75 (6): 555-571,2017.
Pinzone MR, Graf E, Lynch L, McLaughlin B, Hecht FM, Connors M, Migueles SA, Hwang WT, Nunnari G, O'Doherty U: 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 M, Lee ST, Habib J, Robinson C, Silvestri G, O'Doherty U, Chahroudi A: Quantifying integrated SIV-DNA by repetitive-sampling Alu-gag PCR. Journal of Virus Eradication 2 (4): 219-226,2016.
Lee SA, Bacchetti P, Chomont N, Fromentin R, Lewin SR, O'Doherty U, Palmer S, Richman DD, Siliciano JD, Yukl SA, Deeks SG, Burbelo PD: Anti-HIV antibody responses and the HIV reservoir size during antiretroviral therapy. PloS One 11 (8): e0160192,2016.
Imamichi H, Dewar RL, Adelsberger JW, Rehm CA, O'Doherty U, Paxinos EE, Fauci AS, Lane HC: 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 LK, Liu X, VanBelzen DJ, Trinité B, Zheng L, Agosto LM, Migueles SA, Connors M, Sambucetti L, Levy DN, Pasternak AO, O'Doherty U: 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 AO, DeMaster LK, Kootstra NA, Reiss P, O'Doherty U, Berkhout B: Minor contribution of host-HIV readthrough transcripts to the level of HIV cell-associated gag RNA. Journal of Virology 90 (2): 1148-51,2015.
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