Development of a new multi-epitope HIV-1 tat/env/pol/gag vaccine candidate: a preliminary study on immunogenicity

Abstract

Event Abstract Back to Event Development of a new multi-epitope HIV-1 tat/env/pol/gag vaccine candidate: a preliminary study on immunogenicity Roghaye Rahimi1*, Massumeh Ebtekar2*, Neda Arabi3 and Mehdi Mahdavi4 1 Tarbiat Modares University, Iran 2 Tarbiat Modares University, Iran 3 Pasteur institute, Iran 4 Pasteur Institute, Iran Background: Development of new potential vaccine candidates against HIV-1 is highly demanded. Multi-epitope vaccines offer several potential advantages that may be promising in case of mutable divergent pathogens such as HIV-1. In the present study, a multiepitopic recombinant protein containing various HIV-1 antigens was expressed in E. coli BL21D3 and its immunogenicity was initially evaluated in BALB/c mouse. Methods: HIVtop4 sequence spanning the junction of six amino acid fragments (Gag158-186, Pol150-190, ENV296-323, ENV577-610, Tat1-20 and Tat44-61) was designed based on computer analysis to reduce the creation of junctional epitopes, improve the cleavage of proteasome and avoid the accumulation of hydrophobic regions. Synthesized nucleotide sequence corresponding to HIVtop4 was cloned into pET23a plasmid. Expression of pET-HIVtop4 plasmid was induced in BL21 E. coli cells by addition of 1 mM IPTG during 3 hrs culture. Due to C-terminal fusion of 6xHis-tag, the protein was purified by IMAC and further confirmed against anti-His antibody in western-blotting. Groups of BALB/c mice (n=6) were immunized with of 20 g of candidate vaccine adjuvanted in Complete Freund’s adjuvant , Montanide ISI-70 and Alum. Experimental mice groups were immunized three times with 2 weeks interval subcutaneously with 100l of each formulation containing 20 g of vaccine candidate with suitable control groups. Two weeks after last immunization lymphocyte proliferation was measured with Brdu, IL-4 and IFN- cytokine with ELISA, total antibody and IgG1, IgG2a isotypes with indirect ELISA methods. Results: Immunization of mice with HIV-1tat, env, pol, gag led to a significant increase in the proliferative responses of lymphocytes, IFN-γ cytokine and total antibody titer with poly-isotypic form in comparison with the control groups. However Vaccine formulation in alum adjuvant resulted in the highest humoral response and formulation in Freund’s adjuvant resulted in highest level of lymphocyte proliferation and IFN- cytokine release. Conclusion: In this study we concluded tat, env, pol, gag with adjuvants (Montanide, Alum and CFA) can be considered as a candidate vaccine against the HIV virus. Keywords: HIV, Vaccine, Adjuvants, Immunologic, multi epitope vaccine, recombinant protein Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Host-pathogen interactions Citation: Rahimi R, Ebtekar M, Arabi N and Mahdavi M (2013). Development of a new multi-epitope HIV-1 tat/env/pol/gag vaccine candidate: a preliminary study on immunogenicity. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00024 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 09 Mar 2013; Published Online: 22 Aug 2013. * Correspondence: Mrs. Roghaye Rahimi, Tarbiat Modares University, Tehran, Iran, rahimiimmune@gmail.com Dr. Massumeh Ebtekar, Tarbiat Modares University, Tehran, Iran, ebtekarm@modares.ac.ir Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Roghaye Rahimi Massumeh Ebtekar Neda Arabi Mehdi Mahdavi Google Roghaye Rahimi Massumeh Ebtekar Neda Arabi Mehdi Mahdavi Google Scholar Roghaye Rahimi Massumeh Ebtekar Neda Arabi Mehdi Mahdavi PubMed Roghaye Rahimi Massumeh Ebtekar Neda Arabi Mehdi Mahdavi Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.