Clustered epitopes within a new poly-epitopic HIV-1 DNA vaccine shows immunogenicity in BALB/c mice |
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Authors: | Nazli Jafarpour Arash Memarnejadian Mohammad Reza Aghasadeghi Fatemeh Kohram Haniyeh Aghababa Nima Khoramabadi Mehdi Mahdavi |
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Affiliation: | 1. Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran 2. Department of Hepatitis and AIDS, Pasteur Institute of Iran, 69 Pasteur Ave., 1316943551, Tehran, Iran 3. Department of Biotechnology, Tehran Shargh Branch, Payam-e-Nour University, Tehran, Iran 4. Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modarres University, Tehran, Iran 5. Department of Virology, Pasteur Institute of Iran, 69 Pasteur Ave., 1316943551, Tehran, Iran
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Abstract: | Despite a huge number of studies towards vaccine development against human immunodeficiency virus-1, no effective vaccine has been approved yet. Thus, new vaccines should be provided with new formulations. Herein, a new DNA vaccine candidate encoding conserved and immunogenic epitopes from HIV-1 antigens of tat, pol, gag and env is designed and constructed. After bioinformatics analyses to find the best epitopes and their tandem, nucleotide sequence corresponding to the designed multiepitope was synthesized and cloned into pcDNA3.1+ vector. Expression of pcDNA3.1-tat/pol/gag/env plasmid was evaluated in HEK293T cells by RT-PCR and western-blotting. Seven groups of BALB/c mice were intramuscularly immunized three times either with 50, 100, 200 µg of plasmid in 2-week intervals or with similar doses of insert-free plasmid. Two weeks after the last injection, proliferation of T cells and secretion of IL4 and IFN-γ cytokines were evaluated using Brdu and ELISA methods, respectively. Results showed the proper expression of the plasmid in protein and mRNA levels. Moreover, the designed multiepitope plasmid was capable of induction of both proliferation responses as well as IFN-γ and IL-4 cytokine production in a considerable level compared to the control groups. Overall, our primary data warranted further detailed studies on the potency of this vaccine. |
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