Characterization of Intact Proviruses in Blood and Lymph Node from HIV-Infected Individuals Undergoing Analytical Treatment Interruption.

Line K Vibholm,Joy A Pai,Michel C Nussenzweig,Yuria Ablanedo-Terrazas,Julio C C Lorenzi,Marco Garcia Noceda,John P Barton,Thiago Y Oliveira,Tine Damsgaard,Paul W Denton,Perla M Del Rio Estrada,Ching-Lan Lu,Martin Tolstrup,Ole S Søgaard,Yehuda Z Cohen,Gustavo Reyes-Teran

doi:10.1128/jvi.01920-18

Abstract

The role of lymphoid tissue as a potential source of HIV-1 rebound following interruption of antiretroviral therapy (ART) is uncertain. To address this issue, we compared the latent viruses obtained from CD4+ T cells in peripheral blood and lymph nodes to viruses emerging during treatment interruption. Latent viruses were characterized by sequencing near-full-length (NFL) proviral DNA and env from viral outgrowth assays (VOAs). Five HIV-1-infected individuals on ART were studied, four of whom participated in a clinical trial of a TLR9 agonist that included an analytical treatment interruption. We found that 98% of intact or replication-competent clonal sequences overlapped between blood and lymph node. In contrast, there was no overlap between 205 latent reservoir and 125 rebound sequences in the four individuals who underwent treatment interruption. However, rebound viruses could be accounted for by recombination. The data suggest that CD4+ T cells carrying latent viruses circulate between blood and lymphoid tissues in individuals on ART and support the idea that recombination may play a role in the emergence of rebound viremia.IMPORTANCE HIV-1 persists as a latent infection in CD4+ T cells that can be found in lymphoid tissues in infected individuals during ART. However, the importance of this tissue reservoir and its contribution to viral rebound upon ART interruption are not clear. In this study, we sought to compare latent HIV-1 from blood and lymph node CD4+ T cells from five HIV-1-infected individuals. Further, we analyzed the contribution of lymph node viruses to viral rebound. We observed that the frequencies of intact proviruses were the same in blood and lymph node. Moreover, expanded clones of T cells bearing identical proviruses were found in blood and lymph node. These latent reservoir sequences did not appear to be the direct origin of rebound virus. Instead, latent proviruses were found to contribute to the rebound compartment by recombination.

Highlights

The role of lymphoid tissue as a potential source of HIV-1 rebound following interruption of antiretroviral therapy (ART) is uncertain
One potential explanation for this observation is that latent viruses found in CD4ϩ T cells in lymphoid and other tissues differ from those in circulation and that these sequestered cells are responsible for the rebound viremia
To investigate latent viruses from lymph node and blood CD4ϩ T cells, we obtained mononuclear cells from lymph nodes (LNMCs) and peripheral blood (PBMCs) from five HIV-1-infected individuals who had been virally suppressed on ART for a median of 7.3 years (Table 1)

Summary

Introduction

The role of lymphoid tissue as a potential source of HIV-1 rebound following interruption of antiretroviral therapy (ART) is uncertain To address this issue, we compared the latent viruses obtained from CD4ϩ T cells in peripheral blood and lymph nodes to viruses emerging during treatment interruption. To examine the relationship between latent viruses in circulation and in lymph node, we studied five individuals who had concurrent blood draws and lymph node biopsies Four of these individuals were enrolled in a clinical trial that included an analytical treatment interruption (ATI) after 24 weeks of therapy with a TLR9 agonist

Methods

Results

Conclusion