Recombinant HIV type 1 gp120 (ADA) (a.a. 34-518) [His]

As is known, the employment of molecular imprinting polymer (MIP) as specific sensing materials in sensors, namely MIP-based sensors. In this contribution, we devised a MIP electrochemical sensor for the detection of variable-format conformations protein gp120. The sensor was constructed by using a grapheme-like carbon nanfragment (CNF) and bismuth oxides composites (CNF-Bi) as decoration material, small-molecule entry inhibitor NBD-556 and gp120 conjugates NBD-556@gp120 instead of gp120 as the template, and pyrrole as an electropolymerization monomer. Cyclic voltammetry, differential pulse voltammetry, scanning electron microscopy and transmission electron microscope were used to characterize the preparation process of the sensor. Results showing that, under optimized conditions, the introduction of NBD-556 make the specific recognition and analytical properties of the MIP sensor towards gp120 more efficient. The response currents were proportional to the NBD-556@gp120 concentrations in the range of 0.0002 ng mL(-1) to 200 ng mL(-1) with the detection limit of 0.0003 ng mL(-1) based on S/N = 3. Meanwhile, the NBD-556@gp120 based MIP sensor also shows acceptable stability and reproducibility. When used for the detection of gp120 in human plasma, it also showed good accuracy. This research idea is in great promising for the early diagnosis of HIV-1 virus and can also be extended to the detection of other conformationally unstable proteins.

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), Lay 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/mu l), X4 (132 +/- 20 MV/mu l), and Tat (130 +/- 20 MV/ mu l) compared with control (61 +/- 13 MV/mu l). 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 proteinstimulated 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. gpl 20- 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.