Differences in inducibility of the latent HIV reservoir in perinatal and adult infection
Authors: Dhummakupt A, Reubens JH, Anderson T, et al.
Published: Poster presented at 27th Conference on Retroviruses and Opportunistic Infections, March 8th –11th, 2020 – Boston.
Abstract The HIV latent reservoir in resting memory CD4+ T cells prevents cure. Novel therapies to reactivate and eliminate the reservoir are in clinical trials in adults, but not yet in pediatric populations.
HIV proviral reservoir size was determined in perinatal (N=11) and adult infections (N=10) by digital droplet PCR (ddPCR) and with the intact proviral DNA assay (IPDA) in perinatal samples. The inducibility of the latent reservoir was determined with the Tat/rev Induced Limiting Dilution Assay (TILDA) that uses single-round (12 hr) T cell stimulation of CD4+ T cells with PMA/ ionomycin to maximally activate cells to induce proviral expression, measured as multiply-spliced HIV RNA Units per 106 CD4 cells (msRUPM). Markers of immune activation (CD69, CD25 and HLA-DR) and exhaustion (PD-1, TIM-3 and TIGIT) were also assessed. An enhanced TILDA with addition of PHA and for 18 hours was performed to enhance proviral expression in perinatal infections. Non-parametric tests were used for differences between paired and unpaired measurements; correlations were quantified by Spearman rank coefficient.
The median age was 15.8 yrs with a median duration of suppression of 6.7 yrs for perinatal infections, and 40.5 yrs with a median duration of suppression of 7.3 yrs for adult infections. We found that despite a higher proviral reservoir size (median 132.1 vs. 66.7 c/106 PBMCs) and similar rates of T cell activation with PMA/ionomycin (median %CD69 = 96.7% and 93.0%) in perinatal and adult infections, respectively, the size of the induced reservoir was significantly lower in perinatal than in adult infections (median msRUPM of 2.99 vs 11.92, p=0.020). With the enhanced TILDA, the size of the induced reservoir increased significantly in perinatal infections (1.5-fold to a median of 4.5 msRUPM; p=0.034), but not in adult infections. The proportion of induced provirus was significantly lower in perinatal infections at 1.6% compared with 4.0% in adult infections (p=0.030). At baseline, the proportion of HLA-DR+ T cells was significantly lower in perinatal compared with adult infections (median HLA-DR+ cells = 4.56% vs 10.5%, p=0.012), but not correlated with the induced reservoir size.
The inducibility of the latent reservoir is substantially lower in perinatal compared with adult infections, possibly due to differences in baseline states of immune activation, with implications for latency reversal strategies towards ART-free remission.