Fatemeh Tohidi, Seyed Mehdi Sadat*, Azam Bolhassani and Ramin Yaghobi Pages 434 - 439 ( 6 )
Background: Vaccine against HIV-1 is not currently available. In present, Virus like particles (VLPs) as effective strategy was used in several vaccine developing. Two conserved sequences; V3 loop of gp120 and the membrane-proximal external region (MPER) of gp41 are dominant sites for vaccine studies.
Objective: In this study, we used fusion gene of MPER and V3 to product recombinant VLPs and introduced a novel retroviral VLPs harboring high copy of MPER-V3 for HIV-1 vaccine design.
Methods: The pEGFP-N1 plasmid harboring MPER-V3 sequence with Vpr linker was constructed. To produce virus-like particles, HEK 293T cells were co-transfected with the recombinant plasmid, pSPAX-2, pMD2-G and pWPXLd plasmids, evaluated by AFM and SEM microscopy and quantified using P24 end-point ELISA assay.
Results: Time-course quantification of p24 protein as the characteristics of viral production evidenced for the efficient secretion of virus-like structures (up to 120 ng/ml) to the culture supernatant of transfected cells. Examination of the centrifuge-concentrated VLPs by AFM and SEM microscope, also illustrated particles with spherical morphologies and diameters of around 150 nm that had similar sizes to HIV virions.
Conclusion: These data indicated the production of HIV-1 virus-like particles harboring high copy of MPER-V3 that maintained their antigenic structure. These VLPs represented a good implication as a potential vaccine candidate and this guarantees the further investigations towards the assessment of its immunogenicity.
HIV-1, VLP, MPER, V3, recombinant, vaccine.
Department of Microbiology, College of Science, Science and Research Branch, Islamic Azad University, Tehran, Department of Hepatitis, AIDS and Blood- borne Viruses, Pasteur Institute of Iran, Tehran, Department of Hepatitis, AIDS and Blood- borne Viruses, Pasteur Institute of Iran, Tehran, Shiraz Transplant Research Center, Shiraz University of Medical Sciences, Shiraz