Construction and Production of HIV-VLP Harboring MPER-V3 for Potential Vaccine Study.

Abstract:

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.

journal_name

Curr HIV Res

journal_title

Current HIV research

authors

Tohidi F,Sadat SM,Bolhassani A,Yaghobi R

doi

10.2174/1570162X15666171017122229

subject

Has Abstract

pub_date

2017-01-01 00:00:00

pages

434-439

issue

6

eissn

1570-162X

issn

1873-4251

pii

CHR-EPUB-86402

journal_volume

15

pub_type

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