In vitro anti-HIV-1 activity of the recombinant HIV-1 TAT protein along with Tenofovir drug

2020 ◽  
Vol 18 ◽  
Author(s):  
Maryam-Sadat Yadavar-Nikravesh ◽  
Alireza Milani ◽  
Rouhollah Vahabpour ◽  
Mehdi Khoobi ◽  
Haleh Bakhshandeh ◽  
...  
Keyword(s):  
Protein Yield ◽  
Tat Protein ◽  
Hiv Replication ◽  
Toxic Dose ◽  
Expression Host ◽  
Apoptosis Pathways ◽  
Genome Transcription ◽  
Anti Hiv ◽  
Hiv 1

Background: HIV-1 TAT protein is essential for regulation of viral genome transcription. The first exon of TAT protein has a fundamental role in stimulation of the extrinsic and intrinsic apoptosis pathways but its anti-HIV activity is not clear yet. Methods: In the current study, we firstly cloned the first exon of TATcoding sequence in pET-24a expression vector and then protein expression was done in the Rosetta expression host. Next, the expressed TAT protein was purified by Ni-NTA column under native conditions. After that, the protein yield was determined by Bradford kit and NanoDrop spectrophotometry. Finally, the cytotoxicity effect and anti-Scr-HIV-1 activity of the recombinant TAT protein alone and along with Tenofovir drug were assessed by MTT and ELISA, respectively. Results: The recombinant TAT protein was successfully generated in E. coli as confirmed by 13.5% SDS-PAGE and western blotting. The protein yield was ~150-200 µg/ml. In addition, the recombinant TAT protein at a certain dose with low toxicity could suppress Scr-HIV replication in the infected HeLa cells (~30%) that was comparable with a low toxic dose of Tenofovir drug (~40%). It was interesting that the recombinant TAT protein could enhance anti-HIV potency of Tenofovir drug up to 66%. Conclusion: Generally, combination of TAT protein and Tenofovir drug could significantly inhibit HIV-1 replication. It will be required to determine their mechanism of action in the next studies.

scholarly journals In vitro anti-HIV activity of ethanol extract from gandarusa (Justicia gendarussa Burm. f) leaves

2020 ◽  
Author(s):  
Ni Putu Ermi Hikmawanti ◽  
Prihartini Widiyanti ◽  
Bambang Prajogo EW
Keyword(s):  
Ethanol Extract ◽  
Hiv Replication ◽  
Acute Hiv Infection ◽  
Hiv Eradication ◽  
Acute Hiv ◽  
P24 Antigen ◽  
Syncytia Formation ◽  
Anti Hiv ◽  
Hiv 1

Anti retroviral drugs for HIV has problems with uncomfortable side effects and that endanger the lives of HIV sufferers. Several herbs have been empirically proven to have an effect on HIV eradication through inhibition of reverse transcriptase. One of such antiviral herbs is Justicia gendarussa (J. gendarussa). The aim of research is to evaluate anti-HIV activity of 70% fractionated-ethanol extract (with releasing alkaloids) and 70% ethanol extract (without releasing alkaloids) of J. gendarussa leaves on in vitro HIV-infected of MOLT-4 cells. The effect of the extracts in inhibiting viral replication and fusion process on acute HIV infection was identi- fied through syncytia formation assay. Effect of the extracts on HIV p24 antigen was evaluated using HIV-1 p24 ELISA kit. It was found that 70% fractionated-ethanol extract and 70% ethanol extract of J. gendarussa leaves significantly inhibited of HIV replication by inhibition of syncytia formation, where the 50% effective concen- tration (EC50) values of the 70% fractionated-ethanol extract and 70% ethanol extract are 70.5 μg/mL and 228.7 μg/mL, respec- tively. Both of the extracts were also significantly inhibited HIV replication by decreasing HIV p24 antigen level where the EC 50 values of the 70% fractionated-ethanol extract and 70% ethanol extract are 88.8 μg/mL and 540.7 μg/mL, respectively. Moreover, it was found that 70% fractionated-ethanol extract of J. gendarussa leaves has anti-HIV activity since its EC50 values less than 100 μg/mL. It was concluded that J. gendarussa could be potentially developed into a phytopharmaceutical product due to its anti-HIV activity.

Suppression of HIV replication in vitro by CD8+ T-cells from HIV-infected and HIV-seronegative individuals

1997 ◽  
Vol 8 (5) ◽  
pp. 307-310 ◽  
Author(s):  
H Ichimura ◽  
J M Dwyer ◽  
H Tsuchie ◽  
M A Detorio ◽  
M M Hossain ◽  
...  
Keyword(s):  
T Cells ◽  
Nucleotide Sequence ◽  
Cd8 T Cells ◽  
Cd4 T Cells ◽  
Inhibitory Effect ◽  
Hiv Replication ◽  
Anti Hiv ◽  
Hiv 1 ◽  
New Strategies

The inhibitory effect of CD8+ T-cells from HIV-infected or HIVseronegative individuals on HIV replication in the naturally-infected CD4+ T-cells in vitro was examined. Not only autologous CD8+ T-cells from HIV-infected individuals but also allogeneic CD8+ T-cells from HIV-seronegative individuals prevented or delayed HIV replication, even in transwell cocultures using a semipermeable 0.45 micron filter. The level of the inhibitory effect of allogeneic CD8+ Tcells from the HIV-seronegative individuals on the HIV replication was varied among CD4+ T-cells obtained from HIV-infected individuals used. The results suggested that CD8+ T-cells from HIV-seronegative individuals as well as HIVinfected individuals could produce some cytokine(s) which suppress HIV replication in vitro . The sensitivity to the cytokine(s) might be variable among HIV strains, depending on differences in the nucleotide sequence of different HIV-1 strains. Further studies of control of HIV replication by CD8+ anti-HIV cytokine(s) should provide new strategies for the therapy of HIV infection.

Anti HIV-1 Agents 7. Discovery of 1-Hydroxy-4-chloro-9,10-anthraquinone Derivatives as New HIV-1 Inhibitors in Vitro

2011 ◽  
Vol 8 (7) ◽  
pp. 602-605
Author(s):  
Ning Huang ◽  
Qin Wang ◽  
Liu-Meng Yang ◽  
Hui Xu ◽  
Yong-Tang Zheng
Keyword(s):  
Anthraquinone Derivatives ◽  
Anti Hiv ◽  
Hiv 1

scholarly journals Potent Synergistic Anti-Human Immunodeficiency Virus (HIV) Effects Using Combinations of the CCR5 Inhibitor Aplaviroc with Other Anti-HIV Drugs

2008 ◽  
Vol 52 (6) ◽  
pp. 2111-2119 ◽  
Author(s):  
Hirotomo Nakata ◽  
Seth M. Steinberg ◽  
Yasuhiro Koh ◽  
Kenji Maeda ◽  
Yoshikazu Takaoka ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Synergistic Effects ◽  
Immunodeficiency Virus ◽  
Approved Drugs ◽  
Hiv Drugs ◽  
Nonparametric Statistical ◽  
Anti Hiv ◽  
Ccr5 Inhibitor ◽  
Hiv 1

ABSTRACT Aplaviroc (AVC), an experimental CCR5 inhibitor, potently blocks in vitro the infection of R5-tropic human immunodeficiency virus type 1 (R5-HIV-1) at subnanomolar 50% inhibitory concentrations. Although maraviroc is presently clinically available, further studies are required to determine the role of CCR5 inhibitors in combinations with other drugs. Here we determined anti-HIV-1 activity using combinations of AVC with various anti-HIV-1 agents, including four U.S. Food and Drug Administration-approved drugs, two CCR5 inhibitors (TAK779 and SCH-C) and two CXCR4 inhibitors (AMD3100 and TE14011). Combination effects were defined as synergistic or antagonistic when the activity of drug A combined with B was statistically greater or less, respectively, than the additive effects of drugs A and A combined and drugs B and B combined by using the Combo method, described in this paper, which provides (i) a flexible choice of interaction models and (ii) the use of nonparametric statistical methods. Synergistic effects against R5-HIV-1Ba-L and a 50:50 mixture of R5-HIV-1Ba-L and X4-HIV-1ERS104pre (HIV-1Ba-L/104pre) were seen when AVC was combined with zidovudine, nevirapine, indinavir, or enfuvirtide. Mild synergism and additivity were observed when AVC was combined with TAK779 and SCH-C, respectively. We also observed more potent synergism against HIV-1Ba-L/104pre when AVC was combined with AMD3100 or TE14011. The data demonstrate a tendency toward greater synergism with AVC plus either of the two CXCR4 inhibitors compared to the synergism obtained with combinations of AVC and other drugs, suggesting that the development of effective CXCR4 inhibitors may be important for increasing the efficacies of CCR5 inhibitors.

scholarly journals Discovery and Characterization of a Novel CD4-Binding Adnectin with Potent Anti-HIV Activity

2017 ◽  
Vol 61 (8) ◽  
Author(s):  
David Wensel ◽  
Yongnian Sun ◽  
Zhufang Li ◽  
Sharon Zhang ◽  
Caryn Picarillo ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Viral Entry ◽  
High Affinity ◽  
Glycosylation Sites ◽  
Spectrum Activity ◽  
Anti Hiv ◽  
Hiv 1 ◽  
Broad Spectrum Activity

ABSTRACT A novel fibronectin-based protein (Adnectin) HIV-1 inhibitor was generated using in vitro selection. This inhibitor binds to human CD4 with a high affinity (3.9 nM) and inhibits viral entry at a step after CD4 engagement and preceding membrane fusion. The progenitor sequence of this novel inhibitor was selected from a library of trillions of Adnectin variants using mRNA display and then further optimized for improved antiviral and physical properties. The final optimized inhibitor exhibited full potency against a panel of 124 envelope (gp160) proteins spanning 11 subtypes, indicating broad-spectrum activity. Resistance profiling studies showed that this inhibitor required 30 passages (151 days) in culture to acquire sufficient resistance to result in viral titer breakthrough. Resistance mapped to the loss of multiple potential N-linked glycosylation sites in gp120, suggesting that inhibition is due to steric hindrance of CD4-binding-induced conformational changes.

scholarly journals Synthesis and in vitro anti-HIV-1 evaluation of some N-arylsulfonyl-3-formylindoles

2018 ◽  
Vol 54 (3) ◽  
Author(s):  
Zhiping Che ◽  
Yuee Tian ◽  
Shengming Liu ◽  
Mei Hu ◽  
Genqiang Chen
Keyword(s):  
Anti Hiv ◽  
Hiv 1

scholarly journals Mechanism of Action and In Vitro Activity of 1′,3′-Dioxolanylpurine Nucleoside Analogues against Sensitive and Drug-Resistant Human Immunodeficiency Virus Type 1 Variants

1999 ◽  
Vol 43 (10) ◽  
pp. 2376-2382 ◽  
Author(s):  
Zhengxian Gu ◽  
Mark A. Wainberg ◽  
Nghe Nguyen-Ba ◽  
Lucille L’Heureux ◽  
Jean-Marc de Muys ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Mononuclear Cells ◽  
Nucleoside Analogues ◽  
Drug Resistant ◽  
Immunodeficiency Virus ◽  
Blood Mononuclear Cells ◽  
Anti Hiv ◽  
Hiv 1

ABSTRACT (−)-β-d-1′,3′-Dioxolane guanosine (DXG) and 2,6-diaminopurine (DAPD) dioxolanyl nucleoside analogues have been reported to be potent inhibitors of human immunodeficiency virus type 1 (HIV-1). We have recently conducted experiments to more fully characterize their in vitro anti-HIV-1 profiles. Antiviral assays performed in cell culture systems determined that DXG had 50% effective concentrations of 0.046 and 0.085 μM when evaluated against HIV-1IIIB in cord blood mononuclear cells and MT-2 cells, respectively. These values indicate that DXG is approximately equipotent to 2′,3′-dideoxy-3′-thiacytidine (3TC) but 5- to 10-fold less potent than 3′-azido-2′,3′-dideoxythymidine (AZT) in the two cell systems tested. At the same time, DAPD was approximately 5- to 20-fold less active than DXG in the anti-HIV-1 assays. When recombinant or clinical variants of HIV-1 were used to assess the efficacy of the purine nucleoside analogues against drug-resistant HIV-1, it was observed that AZT-resistant virus remained sensitive to DXG and DAPD. Virus harboring a mutation(s) which conferred decreased sensitivity to 3TC, 2′,3′-dideoxyinosine, and 2′,3′-dideoxycytidine, such as a 65R, 74V, or 184V mutation in the viral reverse transcriptase (RT), exhibited a two- to fivefold-decreased susceptibility to DXG or DAPD. When nonnucleoside RT inhibitor-resistant and protease inhibitor-resistant viruses were tested, no change in virus sensitivity to DXG or DAPD was observed. In vitro drug combination assays indicated that DXG had synergistic antiviral effects when used in combination with AZT, 3TC, or nevirapine. In cellular toxicity analyses, DXG and DAPD had 50% cytotoxic concentrations of greater than 500 μM when tested in peripheral blood mononuclear cells and a variety of human tumor and normal cell lines. The triphosphate form of DXG competed with the natural nucleotide substrates and acted as a chain terminator of the nascent DNA. These data suggest that DXG triphosphate may be the active intracellular metabolite, consistent with the mechanism by which other nucleoside analogues inhibit HIV-1 replication. Our results suggest that the use of DXG and DAPD as therapeutic agents for HIV-1 infection should be explored.

scholarly journals In Vitro and In Silico Antioxidant, Anti-Diabetic, Anti-HIV and Anti-Alzheimer Activity of Endophytic Fungi, Cladosporium uredinicola Phytochemicals

2019 ◽  
Vol 13 ◽  
pp. 13-34
Author(s):  
Melappa Govindappa ◽  
V. Thanuja ◽  
S. Tejashree ◽  
C.A. Soukhya ◽  
Suresh Barge ◽  
...  
Keyword(s):  
Qualitative Method ◽  
Cholinesterase Activity ◽  
Methanol Extract ◽  
High Binding Energy ◽  
Acetyl Cholinesterase ◽  
Gp 120 ◽  
Anti Hiv ◽  
Hiv 1

The present work was aimed to identify phytochemicals in C. uredinicola methanol extract from qualitative, TLC and GC-MS method and evaluated for antioxidant, anti-HIV, anti-diabetes, anti-cholinesterase activity in vitro and in silico. The C. uredinicola extract showed flavonoids, tannins, alkaloids, glycosides, phenols, terpenoids, and coumarins presence in qualitative method. From GC-MS analysis, identified seven different phytochemicals and out of seven, four (coumarin, coumarilic acid, hymecromone, alloisoimperatorin) are coumarins. The C. uredinicola extract have shown significant antioxidant activity in DPPH (73) and FRAP (1359) method. The HIV-1 RT (83.81+2.14), gp 120 (80.24+2.31), integrase (79.43+3.14) and protease (77.63+2.14), DPPIV, β-glucosidase and acetyl cholinesterase activity was significantly reduced by the extract. The 2-diphenylmethyleneamino methyl ester had shown significant interaction with oxidant and HIV-1 proteins whereas alloisoimperatorin have interacted with diabetes and cholinesterase proteins followed by hymecromone with high binding energy. These three phytochemicals are non-carcinogens, non-toxic, readily degradable and have drug likeliness properties. The C. uredinicola phytochemicals are responsible for management of diabetes, HIV-1 and Alzheimer. Further in vivo work is needed to justify our research.

scholarly journals Native CGRP Neuropeptide and Its Stable Analogue SAX, But Not CGRP Peptide Fragments, Inhibit Mucosal HIV-1 Transmission

2021 ◽  
Vol 12 ◽  
Author(s):  
Jammy Mariotton ◽  
Anette Sams ◽  
Emmanuel Cohen ◽  
Alexis Sennepin ◽  
Gabriel Siracusano ◽  
...  
Keyword(s):  
T Cell ◽  
Ex Vivo ◽  
Receptor Activation ◽  
Humanized Mice ◽  
Peptide Fragments ◽  
Cgrp Receptor ◽  
Trans Infection ◽  
Anti Hiv ◽  
Hiv 1

BackgroundThe vasodilator neuropeptide calcitonin gene-related peptide (CGRP) plays both detrimental and protective roles in different pathologies. CGRP is also an essential component of the neuro-immune dialogue between nociceptors and mucosal immune cells. We previously discovered that CGRP is endowed with anti-viral activity and strongly inhibits human immunodeficiency virus type 1 (HIV-1) infection, by suppressing Langerhans cells (LCs)-mediated HIV-1 trans-infection in-vitro and mucosal HIV-1 transmission ex-vivo. This inhibition is mediated via activation of the CGRP receptor non-canonical NFκB/STAT4 signaling pathway that induces a variety of cooperative mechanisms. These include CGRP-mediated increase in the expression of the LC-specific pathogen recognition C-type lectin langerin and decrease in LC-T-cell conjugates formation. The clinical utility of CGRP and modalities of CGRP receptor activation, for inhibition of mucosal HIV-1 transmission, remain elusive.MethodsWe tested the capacity of CGRP to inhibit HIV-1 infection in-vivo in humanized mice. We further compared the anti-HIV-1 activities of full-length native CGRP, its metabolically stable analogue SAX, and several CGRP peptide fragments containing its binding C-terminal and activating N-terminal regions. These agonists were evaluated for their capacity to inhibit LCs-mediated HIV-1 trans-infection in-vitro and mucosal HIV-1 transmission in human mucosal tissues ex-vivo.ResultsA single CGRP intravaginal topical treatment of humanized mice, followed by HIV-1 vaginal challenge, transiently restricts the increase in HIV-1 plasma viral loads but maintains long-lasting higher CD4+ T-cell counts. Similarly to CGRP, SAX inhibits LCs-mediated HIV-1 trans-infection in-vitro, but with lower potency. This inhibition is mediated via CGRP receptor activation, leading to increased expression of both langerin and STAT4 in LCs. In contrast, several N-terminal and N+C-terminal bivalent CGRP peptide fragments fail to increase langerin and STAT4, and accordingly lack anti-HIV-1 activities. Finally, like CGRP, treatment of human inner foreskin tissue explants with SAX, followed by polarized inoculation with cell-associated HIV-1, completely blocks formation of LC-T-cell conjugates and HIV-1 infection of T-cells.ConclusionOur results show that CGRP receptor activation by full-length CGRP or SAX is required for efficient inhibition of LCs-mediated mucosal HIV-1 transmission. These findings suggest that formulations containing CGRP, SAX and/or their optimized agonists/analogues could be harnessed for HIV-1 prevention.

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