Methamphetamine activates nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and induces human immunodeficiency virus (HIV) transcription in human microglial cells

Abstract

Human immunodeficiency virus (HIV) primarily infects glial cells in the central nervous system (CNS). Recent evidence suggests that HIV-infected individuals who abuse drugs such as methamphetamine (METH) have higher viral loads and experience more severe neurological complications than HIV-infected individuals who do not abuse drugs. The aim of this study was to determine the effect of METH on HIV expression from the HIV long terminal repeats (LTR) promoter and on an HIV integrated provirus in microglial cells, the primary host cells for HIV in the CNS. Primary human microglial cells immortalized with SV40 T-antigen (CHME-5 cells) were co-transfected with an HIV LTR reporter and the HIV Tat gene, a key regulator of viral replication and gene expression, and exposed to METH. Our results demonstrate that METH treatment induced LTR activation, an effect potentiated in the presence of Tat. We also found that METH increased the nuclear translocation of the nuclear factor kappa B (NF-κB), a key cellular transcriptional regulator of the LTR promoter, and the activity of an NF-κB-specific reporter plasmid in CHME-5 cells. The presence of a dominant-negative regulator of NF-κB blocked METH-related activation of the HIV LTR. Furthermore, treatment of HIV-latently infected CHME-5 (CHME-5/HIV) cells with METH induced HIV expression in a dose-dependent manner, and nuclear translocation of the p65 subunit of NF-κB. These results suggest that METH can stimulate HIV gene expression in microglia cells through activation of the NF-κB signaling pathway. This mechanism may outline the initial biochemical events leading to the observed increased neurodegeneration in HIV-positive individuals who use METH.

Melatonin promotes blood-brain barrier integrity in methamphetamine-induced inflammation in primary rat brain microvascular endothelial cells

“…The nuclear factor-ĸB (NF-κB) signaling is the major pathway that regulates the inflammatory cytokine expression and has been widely studied in inflammatory responses (Kawai and Akira, 2007). Previous studies have reported several stimuli such as lipopolysaccharide (LPS) and METH-induced inflammation in both neuronal cells and glial cells by the overactivation of NF-κB signaling (Flora et al, 2003;Shah et al, 2012;Wires et al, 2012;Perpoonpattana et al, 2013;Coelho-Santos et al, 2015;Jumnongprakhon et al, 2015). Therefore, the inhibition of these negative effects may be beneficial in protecting BBB impairment.…”

RNA-directed gene editing specifically eradicates latent and prevents new HIV-1 infection

“…We inserted DNA oligonucleotides (Table S2) complementary to gRNAs A−D into a humanized Cas9 expression vector (A/B in pX260; C/D in pX330) (16) and tested their individual and combined abilities to alter the integrated HIV-1 genome activity. We first used the microglial cell line CHME5, which harbors integrated copies of a single round HIV-1 vector that includes the 5′ and 3′ LTRs, and a gene encoding an enhanced green fluorescent protein (EGFP) reporter replacing Gag (pNL4-3-ΔGag-d2EGFP) (17). Treating CHME5 cells with trichostatin A (TSA), a histone deacetylase inhibitor, reactivates transcription from the majority of the integrated proviruses and leads to expression of EGFP and the remaining HIV-1 proteome (17).…”

“…We first used the microglial cell line CHME5, which harbors integrated copies of a single round HIV-1 vector that includes the 5′ and 3′ LTRs, and a gene encoding an enhanced green fluorescent protein (EGFP) reporter replacing Gag (pNL4-3-ΔGag-d2EGFP) (17). Treating CHME5 cells with trichostatin A (TSA), a histone deacetylase inhibitor, reactivates transcription from the majority of the integrated proviruses and leads to expression of EGFP and the remaining HIV-1 proteome (17). Expressing of gRNAs plus Cas9 markedly decreased the fraction of TSA-induced EGFPpositive CHME5 cells (Fig.…”

“…TZM-bI reporter and U1 cell lines were obtained from the National Institutes of Health (NIH) AIDS Reagent Program, and CHME5 microglial cells were described previously (17). The detailed procedure for cell growth or preparation of stable cell lines is described in SI Materials and Methods.…”

Effects of HIV-1 Tat and Methamphetamine on Blood-Brain Barrier Integrity and FunctionIn Vitro

“…Experimental and clinical evidence indicates that HIV neuropathogenesis and neurocognitive deficits are exacerbated with coexposure to methamphetamine (30)(31)(32)(33). Methamphetamine induces HIV transcription, potentiates HIV protein-mediated oxidative stress pathways in the brain (34), and increases proinflammatory cytokines (35).…”