Biotechnology Letters

, Volume 41, Issue 11, pp 1283–1298 | Cite as

Design and in vitro delivery of HIV-1 multi-epitope DNA and peptide constructs using novel cell-penetrating peptides

  • Saba Davoodi
  • Azam BolhassaniEmail author
  • Seyed Mehdi Sadat
  • Shiva Irani
Original Research Paper



Developing an effective HIV vaccine that stimulates the humoral and cellular immune responses is still challenging because of the diversity of HIV-1 virus, polymorphism of human HLA and lack of a suitable delivery system.


Using bioinformatics tools, we designed a DNA construct encoding multiple epitopes. These epitopes were highly conserved within prevalent HIV-1 subtypes and interacted with prevalent class I and II HLAs in Iran and the world. The designed DNA construct included Nef60–84, Nef126–144, Vpr34–47, Vpr60–75, Gp16030–53, Gp160308–323 and P248–151 epitopes (i.e., nef-vpr-gp160-p24 DNA) which was cloned into pET-24a(+) and pEGFP-N1 vectors. The recombinant polyepitope peptide (rNef-Vpr-Gp160-P24; ~ 32 kDa) was successfully generated in E. coli expression system. The pEGFP-nef-vpr-gp160-p24 and rNef-Vpr-Gp160-P24 polyepitope peptide were delivered into HEK-293 T cells using cell-penetrating peptides (CPPs). The MPG and HR9 CPPs, as well as the novel LDP-NLS and CyLoP-1 CPPs, were utilized for DNA and peptide delivery into the cells, respectively. SEM results confirmed the formation of stable MPG/pEGFP-N1-nef-vpr-gp160-p24, HR9/pEGFP-N1-nef-vpr-gp160-p24, LDP-NLS/rNef-Vpr-Gp160-P24 and CyLoP-1/rNef-Vpr-Gp160-P24 nanoparticles with a diameter of < 200 nm through non-covalent bonds. MTT assay results indicated that these nanoparticles did not have any major toxicity in vitro. Fluorescence microscopy, flow cytometry and western blot data demonstrated that these CPPs could significantly deliver the DNA and peptide constructs into HEK-293 T cells.


The use of these CPPs can be considered as an approach in HIV vaccine development for in vitro and in vivo delivery of DNA and peptide constructs into mammalian cells.


HIV-1 In silico studies Cell penetrating peptides Vaccine development 


Supporting information

Supplementary Table 1—Top-ranked CABS-dock models of peptide-HLA I complexes.

Supplementary Table 2—Top-ranked CABS-dock models of peptide-HLA II complexes.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

10529_2019_2734_MOESM1_ESM.docx (1.3 mb)
Supplementary file1 (DOCX 1310 kb)
10529_2019_2734_MOESM2_ESM.docx (1.6 mb)
Supplementary file2 (DOCX 1599 kb)


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Saba Davoodi
    • 1
  • Azam Bolhassani
    • 2
    Email author
  • Seyed Mehdi Sadat
    • 2
  • Shiva Irani
    • 1
  1. 1.Department of Biology, School of Basic Science, Science and Research BranchIslamic Azad UniversityTehranIran
  2. 2.Department of Hepatitis and AIDSPasteur Institute of IranTehranIran

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