Persistence of Unintegrated HIV DNA Associates With Ongoing NK Cell Activation and CD34+DNAM-1brightCXCR4+ Precursor Turnover in Vertically Infected Patients Despite Successful Antiretroviral Treatment

Abstract

The quantification of proviral DNA is raising interest in view of clinical management and functional HIV eradication. Measures of all unintegrated HIV DNA (uDNA) forms in infected reservoir cells provides information on recent replication events that is not found from other proviral DNA assays. To evaluate its actual relevance in a cohort of perinatally-infected adult HIV patients (PHIV), we studied how peripheral blood mononuclear cell uDNA levels correlated with total HIV DNA (tDNA) and with overall replication or innate immune control parameters including NK cell activation/exhaustion and lymphoid turnover. Twenty-two PHIV were included, with successfully controlled HIV (HIV RNA <50 copies/mL) on combined antiretroviral therapy for mean of 8.7 ± 3.9 years. uDNA accounted for 16 [5.2-83.5] copies/µg and was strongly correlated with tDNA (ρ=0.700, p=0.001). Flow cytometric analysis of peripheral NK cells showed that CD69 expression was directly correlated uDNA (p=0.0412), but not with tDNA. Interestingly, CD56-CD16+NK cells which include newly described inflammatory precursors and terminally differentiated cells were directly correlated with uDNA levels (p<0.001), but not with tDNA, and an inverse association was observed between the proportion of NKG2D+ NK cells and uDNA (ρ=-0.548, p=0.015). In addition, CD34+DNAM-1brightCXCR4+ inflammatory precursor frequency correlated directly with uDNA levels (ρ=0.579, p=0.0075). The frequencies of CD56-CD16+ and CD34+DNAM-1brightCXCR4+ cells maintained association with uDNA levels in a multivariable analysis (p=0.045 and p=0.168, respectively). Thus, control of HIV-1 reservoir in aviremic patients on ART is an active process associated with continuous NK cell intervention and turnover, even after many years of treatment. Quantification of linear and circular uDNA provides relevant information on the requirement for ongoing innate immune control in addition to ART, on recent replication history and may help stratify patients for functional HIV eradication protocols with targeted options.

CD34+DNAM-1brightCXCR4+ haemopoietic precursors circulate after chemotherapy, seed lung tissue and generate functional innate-like T cells and NK cells

“…The observed increase in CD34 + DNAM -1 bright CXCR4 + cells in PBMC in patients with lung cancer following CT/IT has characteristics of a standard stereotyped defense reaction to inflammatory stimuli (40). This increase is in line with those so far reported during conditions including chronic infection (HIV, HCV) (1, 28), acute infection (SARS-CoV-2) (27), CMV reactivation (10) and persistent/ recurrent inflammation (COPD,PAPA syndrome) (1). Notably, CT/IT uniquely triggered an increase in CD34 + DNAM -1 bright CXCR4 + cells, while conventional CD34 + DNAM-1 -CXCR4and also Lin -CD56 -CD16 + CD7precursors remained unaltered, This observation supports the hypothesis that CD34 + DNAM-1 bright CXCR4 + cells may originate from a distinct mechanism that involves selective recruitment from the bone marrow (BM) in response to CT/IT-induced inflammation.…”

“…We first verified whether CD34 + DNAM-1 bright CXCR4 + “inflammatory” CLP that are detected in PBMC of patients with acute or chronic infections ( 1 , 27 , 28 ) could also be detected in cancer patients. We investigated PBMC of sequential patients on treatment for non-Hodgkin lymphoma, NSCLC, Kaposi’s sarcoma (KS).…”

Primary HIV infection features colonic damage and neutrophil inflammation yet containment of microbial translocation

“…The amount of integrated HIV DNA was obtained by subtracting the amount of uDNA from the amount of total HIV DNA (tDNA). Total/unintegrated/ integrated HIV DNA copy numbers were normalized (as described in [23][24][25][26][27][28][29]) to 1 mg of cellular DNA (equivalent to 142 857 cells) [30].…”