Background:
HIV cure strategies require sophisticated knowledge of the formation and maintenance of latent and active reservoirs of HIV-infected cells. However, the overall paucity of individuals undergoing autopsy have hampered investigation of the anatomical distribution of infected cells during prolonged combination antiretroviral therapy (cART). We report on the largest post mortem study using minimally invasive autopsy on deceased virologically suppressed and unsuppressed adult persons with HIV (PWH) to characterise viral reservoirs.
Methods:
Admitted adult PWH who died at Klerksdorp Tshepong Hospital, North West Province, South Africa were included in this study. Relatives of eligible deceased PWH provided informed consent. Causes of death were documented. Ultrasound-guided core needle biopsies were used to sample a broad range of body fluids and organs ≤16hrs after death. Trocars were used for minimal contamination. Organs were verified by histopathological analyses. HIV reservoir analysis from stored fluid and organ samples was conducted by digital droplet PCR, as HIV copies per million cells (cpm).
Results:
Sixty recently deceased PWH (38 virally suppressed; 22 unsuppressed of whom 13 were cART-naive) were enrolled with a median time to autopsy of 10.4hrs (IQR 6.9). FIND046 was COVID-19 positive and excluded. Most patients were of Black African descent (94.9%), male (57.6%) 50 y/o (IQR 18), with a median VL<24 RNA copies/ml (IQR 2,401). Virally suppressed PWH were on cART for a median of 66.6 months (IQR 80.3). A total of 489 unique specimens from 11 anatomically distinct compartments were analyzed for HIV proviral DNA, including bone marrow aspirate and trephine, brain, bronchioalveolar lavage, cervicovaginal lavage, kidney, lymph nodes, liver, lung, spleen and whole blood. Overall, all anatomical compartments of suppressed and unsuppressed PWH were positive for HIV proviral DNA. Highest copy numbers were detected in lymph nodes (7,384±28,669cpm) and lowest copies in brain (513.7±866cpm) and bone marrow trephine (416±814cpm). No statistically significant differences in HIV proviral DNA concentrations between compartments, irrespective of viremic status were noted. However there was an overall trend of higher proviral DNA in all compartments of viremic patients. Sequence analysis showed viral compartmentalization.
Conclusions:
Findings provide new insights on anatomical location of HIV-infected cells that persist despite prolonged cART that should be targeted for effective cure interventions.