Production and evaluating the properties of HIV-1-Nef-MPER-V3 fusion protein harboring IMT-P8 cell penetrating peptide.

BACKGROUND Finding a safe and effective vaccine against HIV-1 infection is still a major concern and highly valuable. OBJECTIVE This study aimed to design and produce a recombinant Nef-MPER V3 protein fused with IMT-P8, using E. coli expression system to provide a potential HIV vaccine with high cellular penetrance. METHODS After synthesizing the DNA sequence of the fusion protein, the construct was inserted into pET-28 expression vector. The recombinant protein expression was induced using 1 mM IPTG and the product was purified through affinity chromatography. Characterization of cellular delivery, toxicity and immunogenicity of the protein was carried out. RESULTS The recombinant protein was expressed and confirmed by anti-Nef antibody through western blotting. Data analyses showed that the protein has no considerable toxicity effect and has improved the IMT-P8 penetration rate in comparison with a control sample. Moreover, the antigen immunogenicity of the protein induced specific humoral response in mice. CONCLUSION It was concluded that IMT-P8- Nef-MPER-V3 fusion protein has a high penetrance rate into mammalian cell line and low toxicity for potential application as a vaccine against HIV-1.

• Publication year

  2020

• Venue

  Current HIV Research

• Publication date

  2020-06-12

• Fields of study

  Medicine, Chemistry

• Identifiers
  DOI 10.2174/1570162X18666200612151925PMID 32532193
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Comparative analysis of two HIV-1 multiepitope polypeptides for stimulation of immune responses in BALB/c mice.

  2020cited by this paper
• Epidemiologic and viral predictors of antiretroviral drug resistance among persons living with HIV in a large treatment program in Nigeria

  2020cited by this paper
• Production of Recombinant HIV-1 p24-Nef Protein in Two Forms as Potential Candidate Vaccines in Three Vehicles.

  2020cited by this paper
• Effect of HIV Envelope Vaccination on the Subsequent Antibody Response to HIV Infection

  2020cited by this paper
• Induction of a robust humoral response using HIV-1 VLPMPER-V3as a novel candidate vaccine in BALB/c mice.

  2019cited by this paper
• A Novel MVA-Based HIV Vaccine Candidate (MVA-gp145-GPN) Co-Expressing Clade C Membrane-Bound Trimeric gp145 Env and Gag-Induced Virus-Like Particles (VLPs) Triggered Broad and Multifunctional HIV-1-Specific T Cell and Antibody Responses

  2019cited by this paper
• Cell-Penetrating Peptides: Efficient Vectors for Vaccine Delivery

  2019cited by this paper
• Broadly neutralizing antibodies and vaccine design against HIV-1 infection

  2019cited by this paper
• Delivery of HIV-1 polyepitope constructs using cationic and amphipathic cell penetrating peptides into mammalian cells.

  2019cited by this paper
• T cell-based strategies for HIV-1 vaccines

  2019cited by this paper
• How HIV Nef Proteins Hijack Membrane Traffic To Promote Infection

  2019cited by this paper
• Design and in vitro delivery of HIV-1 multi-epitope DNA and peptide constructs using novel cell-penetrating peptides

  2019cited by this paper
• Peptide-Based Vaccination for Antibody Responses Against HIV

  2019cited by this paper
• Cell-penetrating peptides in vaccine delivery: facts, challenges and perspectives.

  2019cited by this paper
• Nanoparticle Vaccines for Inducing HIV-1 Neutralizing Antibodies

  2019cited by this paper
• Multimeric Epitope-Scaffold HIV Vaccines Target V1V2 and Differentially Tune Polyfunctional Antibody Responses.

  2019cited by this paper
• Simultaneous use of natural adjuvants and cell penetrating peptides improves HCV NS3 antigen-specific immune responses.

  2019cited by this paper
• Identification of Immunodominant HIV-1 Epitopes Presented by HLA-C*12:02, a Protective Allele, Using an Immunopeptidomics Approach

  2019cited by this paper
• Updated studies on development of HIV therapeutic vaccine.

  2019cited by this paper
• The breadth of HIV-1 neutralizing antibodies depends on the conservation of key sites in their epitopes

  2019cited by this paper
• A Shot at Dendritic Cell-Based Vaccine Strategy against HIV-1

  2019cited by this paper
• Recombinant HIV-1 vaccine candidates based on replication-defective flavivirus vector

  2019cited by this paper
• Fundamental challenges to the development of a preventive HIV vaccine.

  2018cited by this paper
• In Silico Design and Immunologic Evaluation of HIV-1 p24-Nef Fusion Protein to Approach a Therapeutic Vaccine Candidate

  2018cited by this paper
• HIV Vaccination: A Roadmap among Advancements and Concerns

  2018cited by this paper
• Vaccine Development Strategies, Progresses and Challenges for Human Immunodeficiency Virus (HIV): A Review

  2018cited by this paper
• Life Expectancy after Initiation of Combination Antiretroviral Therapy in Thailand

  2017cited by this paper
• Small heat shock protein 27: An effective adjuvant for enhancement of HIV-1 Nef antigen-specific immunity.

  2017cited by this paper
• Progress in HIV vaccine development

  2017cited by this paper
• The HIV‐1 envelope glycoprotein structure: nailing down a moving target

  2017cited by this paper
• Proteoliposomal formulations of an HIV-1 gp41-based miniprotein elicit a lipid-dependent immunodominant response overlapping the 2F5 binding motif

  2017cited by this paper
• Construction and Production of HIV-VLP Harboring MPER-V3 for Potential Vaccine Study.

  2017cited by this paper
• Combination of cell penetrating peptides and heterologous DNA prime/protein boost strategy enhances immune responses against HIV-1 Nef antigen in BALB/c mouse model.

  2017cited by this paper
• Progress toward active or passive HIV-1 vaccination

  2017cited by this paper
• Conserved HIV Epitopes for an Effective HIV Vaccine

  2017cited by this paper
• Enhancement of viral escape in HIV-1 Nef by STEP vaccination

  2016cited by this paper
• Topical Delivery of Protein and Peptide Using Novel Cell Penetrating Peptide IMT-P8

  2016cited by this paper
• A shot at AIDS.

  2016cited by this paper
• Recent Advances in Lentiviral Vaccines for HIV-1 Infection

  2016cited by this paper
• The structural HCV genes delivered by MPG cell penetrating peptide are directed to enhance immune responses in mice model

  2016cited by this paper
• Vaccine delivery by penetratin: mechanism of antigen presentation by dendritic cells

  2016cited by this paper
• HIV-1 Nef: a master manipulator of the membrane trafficking machinery mediating immune evasion.

  2015cited by this paper
• DNA vaccines: roles against diseases.

  2013cited by this paper
• Plasma IgG to Linear Epitopes in the V2 and V3 Regions of HIV-1 gp120 Correlate with a Reduced Risk of Infection in the RV144 Vaccine Efficacy Trial

  2013cited by this paper
• Broadly neutralizing antibodies against HIV-1: templates for a vaccine.

  2013cited by this paper
• Lessons learned from HIV-1 vaccine trials: new priorities and directions

  2012cited by this paper
• Improving the Endosomal Escape of Cell-Penetrating Peptides and Their Cargos: Strategies and Challenges

  2012cited by this paper
• Application of SCR priming VLP boosting as a novel vaccination strategy against HIV-1.

  2011cited by this paper
• Human immunodeficiency virus (HIV) immunopathogenesis and vaccine development: a review.

  2011cited by this paper
• Induction of HIV Neutralizing Antibodies against the MPER of the HIV Envelope Protein by HA/gp41 Chimeric Protein-Based DNA and VLP Vaccines

  2011cited by this paper
• Membrane topology analysis of HIV-1 envelope glycoprotein gp41

  2010cited by this paper
• Cell-penetrating peptides: application in vaccine delivery.

  2010cited by this paper
• Structure of HIV-1 gp120 with gp41-interactive region reveals layered envelope architecture and basis of conformational mobility

  2009cited by this paper
• Production of recombinant HIV‐1 nef protein using different expression host systems: A techno‐economical comparison

  2009cited by this paper
• Human Immunodeficiency Virus Type 1 Nef Protein Targets CD4 to the Multivesicular Body Pathway

  2009cited by this paper
• High titer HIV-1 V3-specific antibodies with broad reactivity but low neutralizing potency in acute infection and following vaccination.

  2009cited by this paper
• HIV-1 Nef: at the crossroads

  2008cited by this paper
• Role of HIV Gp41 mediated fusion/hemifusion in bystander apoptosis

  2008cited by this paper
• V3: HIV's Switch-Hitter

  2005cited by this paper
• Anti-Human Immunodeficiency Virus Type 1 (HIV-1) Antibodies 2F5 and 4E10 Require Surprisingly Few Crucial Residues in the Membrane-Proximal External Region of Glycoprotein gp41 To Neutralize HIV-1

  2005cited by this paper
• CD8+ T Cell Epitope-Flanking Mutations Disrupt Proteasomal Processing of HIV-1 Nef1

  2005cited by this paper
• HIV vaccine design and the neutralizing antibody problem

  2004cited by this paper
• Protein cargo delivery properties of cell-penetrating peptides. A comparative study.

  2004cited by this paper
• Epitope-based vaccines: an update on epitope identification, vaccine design and delivery.

  2003cited by this paper
• Epitope-vaccine strategy against HIV-1: today and tomorrow.

  2003cited by this paper
• Two-step purification of His-tagged Nef protein in native condition using heparin and immobilized metal ion affinity chromatographies.

  2003cited by this paper
• Viral persistence: HIV's strategies of immune system evasion.

  2002cited by this paper
• Structure–function relationships in HIV‐1 Nef

  2001cited by this paper
• Recombinant nef HIV-IIIB protein is toxic to human neurons in culture.

  2000cited by this paper

• Comparative Analysis of IMT-P8 and LDP12 Cell-Penetrating Peptides in Increasing Immunostimulatory Properties of HIV-1 Nef-MPER-V3 Antigen.

  2024cites this paper