Measurement for Viral Reservoir and Immune Function in HIV-1-infected Patients Under Antiretroviral Therapy
Condition: HIV-1-infection
Study type: Observational

Summary: 
      Current antiretroviral therapy (ART) is highly effective to suppress plasma viral load to
      below the detection limit and to restore the host immunity, thus to prolong the survival of
      HIV-1-infected patients remarkably. However, HIV-1 will rebound to pre-treatment levels
      within weeks of interruption or irregular medication. The reason why HIV-1 would not be
      eradicated by powerful ART can be explained by that the reservoir of latent HIV-1 in resting
      CD4 T cells will persistently exist even long-term suppression of plasma viral RNA. Several
      therapeutic approaches that aim to prevent or delay viral rebound after treatment
      interruption, producing a post-treatment remission or functional cure of HIV-1, are being
      investigated. This study is to measure the size of viral reservoir and HIV-1-specific T cell
      response in HIV-1-infected patients during ART to help understand the mechanism of HIV-1
      persistence, then to help establish a potential policy for functional cure.
    

Detailed description: 
      Introduction

      Current antiretroviral therapy (ART) is highly effective to suppress plasma viral load to
      below the detection limit and to restore the host immunity, thus to prolong the survival of
      HIV-1-infected patients remarkably. However, HIV-1 will rebound to pre-treatment levels
      within weeks of interruption or irregular medication1,2. The reason why HIV-1 would not be
      eradicated by powerful ART can be explained by that the reservoir of latent HIV-1 in resting
      CD4 T cells will persistently exist even long-term suppression of plasma viral RNA. Several
      therapeutic approaches that aim to prevent or delay viral rebound after treatment
      interruption, producing a post-treatment remission or functional cure of HIV-1, are being
      investigated3. This study is to measure the size of viral reservoir and HIV-1-specific T cell
      response in HIV-1-infected patients during ART to help understand the mechanism of HIV-1
      persistence, then to help establish a potential policy for functional cure.

      Methods Enrollment criteria

        1. Confirmed HIV-1 infected patients under regular follow-up and receiving antiretroviral
           therapy

        2. Men with 20-50 years old

      Exclusion criteria

        1. Serious co-morbidity

        2. Obvious hepatic or renal dysfunction

        3. Receiving immunosuppressive therapy

      Duration and frequency A total of 48 weeks of follow-up, and laboratory measurement every 8
      weeks

      Laboratory measurement We will draw the blood specimen from subjects after they sign the
      informed consent. About 30 mL blood will be taken and sent for the determination of viral
      reservoir and HIV-1-specific T cell responses, every 8 weeks, from the baseline at enrollment
      to the end of 48 weeks.

      HIV-1 Gag-specific T-cell responses Recombinant HIV-1 Gag p24 (5 ug/mL; Research Diagnostics,
      Inc.) will be used as HIV-1 antigen. PBMC (106) will be incubated with or without p24 in
      24-well plats for 6 days. The frequency of T cells with BrdU incorporation will be measured
      by flow cytometry.

      HIV Viral Reservoirs

      Levels and contents of HIV viral reservoirs in the peripheral blood will be analyzed in CD4+
      T cells(the major cell type harboring HIV reservoir) in 2 major components:

        -  Proviral DNA

        -  Cell-associated (CA) HIV RNAs The quantifications for both HIV pvDNA and CA RNAare based
           on the PCRbasedquantitative system.

      Proviral DNA Assessment The blood samples will be used to isolate CD4+ T cells for cellular
      DNA extraction.

      Standardized aliquot amount of extracted DNA is used to determine HIV pvDNA level
      throughPCR-based quantitative assay targeting the conserved region of HIV gene. The HIV
      pvDNAlevel will be presented as copies per million CD4+ T cells.The input cell number is
      determined by a separate PCR targeting human endogenous gene(s).

      Cell-associated (CA) HIV RNA Total cellular RNA will be extracted from CD4+ T cells to
      generate totalcomplementary DNA (cDNA) through reverse transcription. The cDNA is then used
      to determine CA HIV RNA level through PCR-based quantitative assay that targets the conserved
      sequences of HIV at a designated laboratory by Sponsor. The CA RNA level is presented a
      scopies of endogenous house-keeping gene transcripts, whose copy number will be determined
      independently by a separate PCR targeting the RNA transcripts of the house-keeping gene.
    

Intervention name(s): no intervention
Intervention type(s): Other
Intervention description(s): No intervention
Observational model: Cohort
Time perspective: Prospective
Sampling method: Non-Probability Sample
Study population: 
        HIV-1 infected patients receiving antiretroviral treatment
      
Biospecimen description: 
      sample with DNA