Abstract:
OBJECTIVES:A potent HIV vaccine should overcome some limitations such as polymorphism of human HLA, the diversity of HIV-1 virus, and the lack of an effective delivery system. In this study, a DNA construct encoding Nef60-84, Nef126-144, Vpr34-47, Vpr60-75, Gp16030-53, Gp160308-323, and P248-151 epitopes was designed using bioinformatics tools. The pcDNA3.1-nef-vpr-gp160-p24 and pcDNA3.1-nef constructs were prepared in large scale as endotoxin-free form. Moreover, the recombinant Nef-Vpr-Gp160-p24 polypeptide and Nef protein were generated inE. coli. These constructs were delivered using cell penetrating peptides (CPPs) in vivo, and immune responses were assessed for different modalities in BALB/c mice. RESULTS:The recombinant DNA constructs were confirmed as the ~ 867 bp and ~ 648 bp bands related tonef-vpr-gp160-p24 andnef genes on agarose gel. Moreover, the purified Nef-Vpr-Gp160-p24 polypeptide and Nef protein showed the ~ 32 kDa and ~ 30 kDa bands on SDS-PAGE, respectively. The results of immune responses indicated that the heterologous prime/boost regimens using both Nef-Vpr-Gp160-P24 and Nef antigens induced significantly the secretion of IgG2a, IgG2b, IFN-γ and Granzyme B compared to other groups. The levels of Granzyme B in mice immunized with Nef antigen were higher than those immunized with Nef-Vpr-Gp160-P24 antigen. The CPPs showed the same potency with Montanide adjuvant for eliciting immune responses. CONCLUSIONS:The heterologous prime/boost regimens for both antigens could significantly direct immune responses toward Th1 and CTL activity compared to other regimens. Comparing the efficiency of Nef-Vpr-Gp160-P24 and Nef constructs, the Nef-Vpr-Gp160-P24 constructs delivered by CPPs showed promising results as an HIV vaccine candidate.
journal_name
Biotechnol Lettjournal_title
Biotechnology lettersauthors
Davoodi S,Bolhassani A,Namazi Fdoi
10.1007/s10529-020-03060-3subject
Has Abstractpub_date
2021-01-01 00:00:00eissn
0141-5492issn
1573-6776pii
10.1007/s10529-020-03060-3pub_type
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journal_title:Biotechnology letters
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doi:10.1007/s10529-006-9020-z
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pub_type: 杂志文章
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doi:10.1007/s10529-010-0227-7
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journal_title:Biotechnology letters
pub_type: 杂志文章,评审
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pub_type: 杂志文章,评审
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pub_type: 杂志文章
doi:10.1023/a:1023402529882
更新日期:2003-05-01 00:00:00
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journal_title:Biotechnology letters
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更新日期:2011-03-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-016-2237-6
更新日期:2017-02-01 00:00:00
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pub_type: 杂志文章
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更新日期:2016-03-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-019-02696-0
更新日期:2019-09-01 00:00:00
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pub_type: 杂志文章
doi:10.1007/s10529-017-2394-2
更新日期:2017-12-01 00:00:00
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pub_type: 杂志文章
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更新日期:2009-02-01 00:00:00