Little is known about autophagy during HIV infection. Recently, two different teams reported important dysfunctions of autophagy in HIV-infected patients despite sustained suppressive antiretroviral therapy. As altered autophagy is strongly linked to cellular senescence and chronic inflammation, two hallmarks of HIV-infected patients despite long-term suppressive antiretroviral therapy, it is important to improve our knowledge in the area.
Our main objective is to determine whether all or part of mononuclear cell subpopulations (CD4+ and CD8+ T lymphocytes, and monocytes) exhibit a defect in autophagy function in a cohort of HIV-infected patients who are virologically-controlled (plasma HIV RNA \<50 copies / ml) either spontaneously (i.e. HIV controllers or post-treatment controllers) or after they started antiretroviral therapy at different time points (i.e. at the acute or chronic phases), as compared with a control group (i.e. uninfected healthy blood donors).
Age >=18 years
Man or woman
Infected with HIV-1 (and not co-infected with HIV-2)
Followed at Orleans' Regional Hospital
Patient unable, according to the investigator, to meet the requirements of the protocol
Pregnant or lactating woman
Patient with a history of inflammatory bowel disease, malignancy, intestinal ischemia, malabsorption or other gastrointestinal dysfunction that, in the judgment of the investigator, could interfere with the interpretation of the results.
Presence of coagulation abnormality or unexplained bleeding history
All HIV-1-infected adults (not co-infected with HIV-2), followed at the Infectious Diseases Department of Orleans' Regional Hospital and meeting the inclusion criteria, will be offered to participate in the study.
Cohort
Cross-sectional
expression of a panel
Biological/VaccineQuantify, by droplet digital PCR, the expression of a panel of 7 genes (+ GALIG) involved in autophagy2 on sub-populations (CD4+ and CD8+ lymphocytes and monocytes) after their sorting using magnetic bead cell then RNA extraction Evaluate, on a functional test (as previously described1), whether the observed expression dysregulation is associated with a deregulation of the autophagic function, whether constitutive or induced.
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Treatment groupsSee each treatment group below.
Quantify of a panel of genes involved in autophagy on sub-populations
Quantify by droplet digital PCR, the expression of a panel of 7 genes (+ GALIG) involved in autophagy on sub-populations (CD4+ and CD8+ lymphocytes and monocytes) after their sorting using magnetic bead cell then RNA extraction
functional test on mononuclear cell subpopulations in the autophagy function
Evaluate on a functional test whether the observed expression dysregulation is associated with a deregulation of the autophagic function, whether constitutive or induced.
Validation of the expression assays of genes involved in the autophagy function
Validate the expression assays of genes involved in the autophagy process and the statistical analyzes obtained on PBMCs on a validation cohort.
analyze regulatory markers on dysregulated gene promoters
To analyze regulatory markers on dysregulated gene promoters, we will analyze and compare the epigenetic marks (acetylation and methylation of histones, DNA methylation) overall and on the promoter zones of autophagy genes which present an alteration of expression.
quantify the reservoir of virus and ongoing viral replication
To quantify the reservoir of virus and ongoing viral replication, we will respectively evaluate the integrated viral DNA and the 2LTR circles levels.
analyze the phenotype of the PBMCs studied
To analyze the phenotype of the PBMCs studied, whether HIV positive or negative, we will evaluate, by analysis of surface markers in flow cytometry: * Distribution of CD3+CD4+, CD3+CD8+, NK, B, NK-T and monocyte populations * Activation (HLA-DR and CD38 on T cells; HLA-DR, CD11b and CD16 on monocytes) * Senescence (CD57+ on CD8+ T cells, CD7- on CD4+ T cells) * Exhaustion (PD1 / CD279) * Distribution of memories, naive and effector CD4+ and CD8+ lymphocytes (CD45RA and CCR7 / CD197) * Proliferative capacity of PBMCs
Evaluation of the level of inflammation
To assess the level of inflammation, we will dose the inflammatory cytokines present in the patients' serum and controls.
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