Effects of HIV-1 gp120 and TAT-derived microvesicles on endothelial cell function

Abstract

The aims of this study were twofold. The first was to determine if human immunodeficiency virus (HIV)-1 glycoprotein (gp) 120 and transactivator of transcription (Tat) stimulate the release of endothelial microvesicles (EMVs). The second was to determine whether viral protein-induced EMVs are deleterious to endothelial cell function (inducing endothelial cell inflammation, oxidative stress, senescence and increasing apoptotic susceptibility). Human aortic endothelial cells (HAECs) were treated with recombinant HIV-1 proteins Bal gp120 (R5), Lav gp120 (X4), or Tat. EMVs released in response to each viral protein were isolated and quantified. Fresh HAECs were treated with EMVs generated under control conditions and from each of the viral protein conditions for 24 h. EMV release was higher ( P < 0.05) in HAECs treated with R5 (141 ± 21 MV/µl),X4 (132 ± 20 MV/µl), and Tat (130 ± 20 MV/µl) compared with control (61 ± 13 MV/µl). Viral protein EMVs induced significantly higher endothelial cell release of proinflammatory cytokines and expression of cell adhesion molecules than control. Reactive oxygen species production was more pronounced ( P < 0.05) in the R5-, X4- and Tat-EMV-treated cells. In addition, viral protein-stimulated EMVs significantly augmented endothelial cell senescence and apoptotic susceptibility. Concomitant with these functional changes, viral protein-stimulated EMVs disrupted cell expression of micro-RNAs 34a, 126, 146a, 181b, 221, and miR-Let-7a ( P < 0.05). These results demonstrate that HIV-1 gp120 and Tat stimulate microvesicle release from endothelial cells, and these microvesicles confer pathological effects on endothelial cells by inducing inflammation, oxidative stress, and senescence as well as enhancing susceptibility to apoptosis. Viral protein-generated EMVs may contribute to the increased risk of vascular disease in patients with HIV-1.NEW & NOTEWORTHY Human immunodeficiency virus (HIV)-1-related proteins glycoprotein (gp) 120 and transactivator of transcription (Tat)-mediated endothelial damage and dysfunction are poorly understood. Endothelial microvesicles (EMVs) serve as indicators and potent mediators of endothelial dysfunction. In the present study we determined if HIV-1 R5- and X4-tropic gp120 and Tat stimulate EMV release in vitro and if viral protein-induced EMVs are deleterious to endothelial cell function. gp120 and Tat induced a marked increase in EMV release. Viral protein-induced EMVs significantly increased endothelial cell inflammation, oxidative stress, senescence, and apoptotic susceptibility in vitro. gp120- and Tat-derived EMVs promote a proinflammatory, pro-oxidative, prosenescent, and proapoptotic endothelial phenotype and may contribute to the endothelial damage and dysfunction associated with gp120 and Tat.

Circulating Cell Biomarkers in Pulmonary Arterial Hypertension: Relationship with Clinical Heterogeneity and Therapeutic Response

“…Recent studies show that EMVs, identified as CD62E + , are significantly increased in patients with HIV infection treated with antiretroviral therapy when compared to controls [ 36 ]. Additionally, Hijmans et al showed that HIV-1 proteins are able to induce EMV release from endothelial cells and to promote endothelial senescence, inflammation and oxidative stress [ 37 ].…”

Transient Hyperglycemia and Hypoxia Induce Memory Effects in AngiomiR Expression Profiles of Feto-Placental Endothelial Cells

“…PCR efficiency of all primer assays was tested using a five-point standard curve with a range from 25 to 0.04 ng/reaction of pooled cDNA in triplicates. Expression of angiomiRs was relatively quantified (2 −∆∆Ct method) [57] using two distinct strategies. The expression of miR-199a-3p was very low and below the detection limit in 14% of samples, and thus, was excluded from the analysis.…”

Unconventional Pathways of Protein Secretion: Mammals vs. Plants

“…Like the blebbing mechanism, firstly the cytoskeleton adjacent to the site of shedding on the plasma membrane is disassembled, then the phosphatidylserine is translocated to the outer leaflet causing the plasma membrane to bulge ( Cohen et al, 2020 ). Functional microvesicles are involved in several physiopathologic conditions, such as inflammation, oxidative stress, and senescence ( Hijmans et al, 19852019 ). These vesicles differ from exosomes not only in formation mechanism, but also in size and molecular markers.…”