Bystander cell death by pyroptosis of astrocytes requires cell-to-cell transmission of HIV

OJEDA DIEGO; CEVALLOS CINTIA; URQUIZA JAVIER; QUARLERI JORGE; SANCHEZ LAUTARO; LAUFER NATALIA

Mexico

Conferencia; 10th IAS Conference on HIV Science (IAS 2019); 2019

IAS

The human immunodeficiency virus (HIV) is capable of infecting the brain parenchyma, producing neurological disorders associated with inflammation in the central nervous system (CNS), which persists despite combined antiretroviral treatment. Astrocytes, the most abundant cell type in the CNS, play a leading role in neuropathogenesis, promoting neuronal damage during viral infections of the CNS. The mechanisms involved in astrocytic pathogenesis remain obscure.Normal human astrocytes (NHA) were infected in vitro using pseudotyped HIV-VSV-G co-expressing the green fluorescent protein (GFP). The infection was monitored by flow cytometry (FACS), discriminating in each culture productively infected cells (PI / GFP +) from those that were not (NPI / GFP-), and simultaneously, mitochondrial membrane potential (ΔΨm) and cell death (AnnexinV+/ 7AAD+) were evaluated. PI and NPI cells were sorted by FACS in order to measure pyroptosis differentially (NLRP3, cleaved forms of caspase-1 and GSDMD by WB). Besides, the role of PI and NPI on bystander cellular damage was measured in co-culture assays using vital proliferation dye (VCP) and FACS. To infer the viral stimuli associated with the activation of the pyroptosis, antiviral drugs (zidovudina, nevirapine, raltegravir and indinavir) were used to inhibit viral enzymes involved in the replication.NPI cells (but not PIs) exhibited significantly higher ΔΨm dissipation and cell death (p