In vitro Delivery of HIV-۱ Nef Antigen by Histidine-rich nona-arginine and Latarcin ۱ peptide

Publish Year: 1398
نوع سند: مقاله ژورنالی
زبان: English
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JR_JMMI-7-4_003

تاریخ نمایه سازی: 29 بهمن 1402

Abstract:

Introduction: The Nef accessory protein is an attractive antigenic candidate in the development of HIV-۱ DNA- or protein-based vaccines. The most crucial disadvantage of DNA and protein-based vaccines is their low immunogenicity, which can be improved by cell-penetrating peptides (CPPs) as effective carrier molecules. Methods: In this study, the HIV-۱ Nef protein was generated in the Escherichia coli expression system for in vitro delivery using a novel CPP, Latarcin ۱ peptide, in a non-covalent manner. Also, the Histidine-rich nona-arginine peptide was utilized to transfer the HIV-۱ Nef gene. The size, morphology, and zeta potential of the complexes were evaluated by scanning electron microscopy (SEM) and Zetasizer. The efficiency of cell transfection was studied using a fluorescence microscopy and flow cytometry for the DNA/CPP complexes and western blot analysis for the protein/CPP complexes. Results: The Nef protein generated in the BL۲۱ strain migrated as a dominant band of ~۳۰ kDa in SDS-PAGE. The SEM data confirmed the formation of stable complexes with a size below ۲۰۰ nm. MTT assay demonstrated that the complexes did not represent any considerable cytotoxic effect compared to untreated HEK-۲۹۳T cells. The results of fluorescence microscopy, flow cytometry, and western blotting revealed that the Nef DNA and protein constructs could be significantly transfected into HEK-۲۹۳T cell line using these CPPs. Conclusion: These data suggest that the Histidine-rich nona-arginine peptide and Latarcin ۱ peptide as CPPs can be considered as a promising approach in the development of the HIV-۱ vaccine for gene or protein delivery.

Authors

Fatemeh Namazi

Department of Biology, School of Basic Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran

Azam Bolhassani

Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran

Seyed Mehdi Sadat

Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran

Shiva Irani

Department of Biology, School of Basic Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran

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