Design and in vitro delivery of HIV-1 multi-epitope DNA and peptide constructs using novel cell-penetrating peptides

Davoodi, Saba; Bolhassani, Azam; Sadat, Seyed Mehdi; Irani, Shiva

doi:10.1007/s10529-019-02734-x

Cited by 20 publications

(9 citation statements)

References 51 publications

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“…This is mainly attributed to the main role of Nef in HIV pathogenesis via downregulation of cell-surface expression of multiple membrane-associated proteins, enhancement of virion infectivity, stimulation of viral replication, interference of cellular 15/ Nikyar and colleagues polymerase. The pET vectors have been widely used for designing HIV proteins expression constructs (16,(25)(26)(27). In the present study, we successfully constructed a recombinant fusion plasmid encoding a chimeric Nef-Vpr gene in pET23a expression vector for generation of a recombinant fusion protein in near future.…”

Section: Discussionmentioning

confidence: 97%

Construction of a Prokaryotic Expression Vector harboring Two HIV-1 Accessory Genes

et al. 2021

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Background and objectives: HIV-1 Nef and Vpr antigens have been described as suitable candidates for therapeutic HIV vaccine development. The aim of this study was to generate Nef-Vpr fusion gene construct and to clone the construct into pET-23a (+), a prokaryotic expression vector.Methods: HIV-1 Nef and Vpr genes were PCR-amplified from the pNL4-3 plasmid using specific primers and Pfu DNA polymerase. Results of PCR amplification were visualized by electrophoresis on 0.8% agarose gel. At first, the amplified Nef fragment was cloned into NheI and BamHI restriction sites of pET-23a expression vector. Next, cloning of Vpr gene was performed into BamHI and HindIII restriction sites of the pET-23a-Nef vector. Finally, purity of the recombinant pET-23-Nef-Vpr construct was determined by NanoDrop spectrophotometry.Results: PCR amplification of Nef and Vpr genes was confirmed by detection of 620 bp and 291 bp bands, respectively. Cloning of the Nef-Vpr construct into the vector was confirmed by detection of a 911 bp fragment following enzymatic digestion with NheI and HindIII and sequencing. Conclusion:The successful construction of recombinant fusion plasmid encoding a chimeric Nef-Vpr gene was performed in a prokaryotic expression vector for development of HIV-1 recombinant protein vaccine in near future.

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Section: Discussionmentioning

confidence: 97%

Construction of a Prokaryotic Expression Vector harboring Two HIV-1 Accessory Genes

et al. 2021

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“…Epitope Conservancy Analysis among HIV subtypes and population coverage was performed using the IEDB website. The interaction between the selected epitopes and MHCs was studied by the peptide-protein docking websites (3,(17)(18)(19). Finally, the peptide constructs were designed using the selected epitopes.…”

Section: Methodsmentioning

confidence: 99%

“…The high genetic variability of HIV-1 has led to some problems in the prevention, diagnosis, and treatment of AIDS [2]. Thus, scientists are trying to determine the conserved and immunogenic epitopes using bioinformatics tools in all populations to develop HIV therapeutic vaccine and diagnosis [3]. At present, HIV is diagnosed by various immunological and molecular techniques.…”

Section: Introductionmentioning

confidence: 99%

Detection of Anti-IgGs against Heat Shock Proteins 27 and 20, HP91 Peptide, and HIV-1 Polypeptides in HIV-Positive and Negative Patients

Milani

Baesi

Agi

et al. 2020

JoMMID

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A simple and sensitive diagnosis method is needed to identify HIV infection in sera of untreated, treated, and drug-resistant patients. The purpose of this study is to determine whether heat shock proteins (Hsp)-27 and -20 and HP91 peptide along with HIV-1 polypeptides can serve as potential biomarkers to distinguish HIV infection in untreated, treated, and drug-resistant individuals compared to HIV-negative subjects. Methods: At first, human sera were obtained from 141 participants, including 20 naïve HIV-infected, 71 treated, 30 drugresistant, 20 HIV-negative (healthy/control) individuals. The recombinant Hsp27, Hsp20, and five designed HIV-1 polypeptides were expressed in Escherichia coli and purified by affinity chromatography under denaturing or native conditions. Finally, the antibodies against these antigens were quantified in sera using ELISA. Results: Our data showed that HIV-infected patients significantly displayed higher serum levels of anti-Hsp27, anti-HP91, and anti-Nef-Tat-Gp160-P24, anti-Nef-Vpr-Gp160-P24, anti-Nef-Vif-Gp160-P24, anti-Nef-Vpu-Gp160-P24, and anti-Nef-Rev-Gp160-P24 polypeptide antibodies than healthy groups (p < 0.05), but not for anti-Hsp20. Moreover, the serum levels of antibodies against Hsp27, Hsp20, HP91, and HIV-1 polypeptides were not statistically significant between different groups of patients (p > 0.05). Conclusion: The levels of anti-Hsp27 and anti-HP91 antibodies in serum increased in HIV-1 seropositive subjects along with antibodies against five HIV-1 polypeptides suggesting their potential value as a diagnostic marker for HIV-1 infections.

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“…The designed DNA included genes for nef-vpr-gp160-p24 epitopes and was complexed with MPG through noncovalent interactions. The complexes were able to interact with cells and induce humoral and cellular immune responses in vivo [264]. MPG was also used as a delivery system for the hepatitis C virus (HCV) antigens.…”

Section: Cell-penetrating Peptidesmentioning

confidence: 99%

Use of Protamine in Nanopharmaceuticals—A Review

et al. 2021

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Macromolecular biomolecules are currently dethroning classical small molecule therapeutics because of their improved targeting and delivery properties. Protamine-a small polycationic peptides represent a promising candidate. In nature, it binds and protects DNA against degradation during spermatogenesis due to electrostatic interactions between the negatively charged DNA-phosphate backbone and the positively charged protamine. Researchers are mimicking this technique to develop innovative nanopharmaceutical drug delivery systems, incorporating protamine as a carrier for biologically active components such as DNA or RNA. The first part of this review highlights ongoing investigations in the field of protamine-associated nanotechnology, discussing the self-assembling manufacturing process and nanoparticle engineering. Immune-modulating properties of protamine are those that lead to the second key part, which is protamine in novel vaccine technologies. Protamine-based RNA delivery systems in vaccines (some belong to the new class of mRNA-vaccines) against infectious disease and their use in cancer treatment are reviewed, and we provide an update on the current state of latest developments with protamine as pharmaceutical excipient for vaccines.

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Design and in vitro delivery of HIV-1 multi-epitope DNA and peptide constructs using novel cell-penetrating peptides

Cited by 20 publications

References 51 publications

Construction of a Prokaryotic Expression Vector harboring Two HIV-1 Accessory Genes

Construction of a Prokaryotic Expression Vector harboring Two HIV-1 Accessory Genes

Detection of Anti-IgGs against Heat Shock Proteins 27 and 20, HP91 Peptide, and HIV-1 Polypeptides in HIV-Positive and Negative Patients

Use of Protamine in Nanopharmaceuticals—A Review

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