Cloning, Expression and Purification of a Novel Multi-epitopic HIV-1 Vaccine Candidate: A Preliminary Study on Immunoreactivity

Authors

  • Arash Memarnejadian Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran
  • Fatemeh Kohram Department of Biotechnology, Tehran Shargh Branch, Payam-e-Nour University, Tehran, Iran
  • Haniyeh Aghababa Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University
  • Mehdi Mahdavi Department of Immunology, Pasteur Institute of Iran, Tehran, Iran
  • Mohammad Reza Aghasadeghi Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran
  • Morteza Taghizadeh Department of Medical virology, Tehran University of Medical Science, Tehran, Iran
  • Nima Khoramabadi Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University
  • Samira Arabi Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
  • Zahra Shahosseini Department of Immunology, Pasteur Institute of Iran, Tehran, Iran
Abstract:

  Introduction : Designing an effective vaccine against human immunodeficiency virus (HIV)-1 is a global health priority . Multi-epitope vaccines offer several potential advantages that may be promising in case of mutable divergent pathogens such as HIV-1. Herein, a multiepitopic recombinant protein containing various HIV-1 antigens was expressed in E. coli cells and its immunogenicity in combination with different adjuvants 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 immunoinformatic analysis to reduce the creation of junctional epitopes, improve the cleavage of proteasome and avoid the local accumulation of hydrophobic regions. Synthesized nucleotide sequence corresponding to HIVtop4 was cloned into pET23a plasmid. Expression of pET-HIVtop4 plasmid was induced in BL21 (DE3) E. coli cells by addition of 1 mM IPTG during 3 h culture and the protein was purified by Ni-NTA column chromatography and further confirmed against anti-His antibody in western-blotting. Groups of BALB/c mice (n=6) were immunized three times with 2 weeks interval, subcutaneously with 10 m g of candidate vaccine adjuvanted in Complete Freund’s adjuvant , Montanide ISA70 and Alum with suitable control groups. Two weeks after last immunization lymphocyte proliferation was measured with Brdu, IL-4 and IFN- g cytokines with ELISA, total antibody and IgG1, IgG2a isotypes with indirect ELISA methods. Results: Results showed that Immunization with HIV-1 tat/pol/gag/env led to a significant increase in the proliferative responses of lymphocytes, IL-4 and IFN-γ cytokine production and humoral immune response in comparison with the control groups. Conclusion: In this study we concluded that Tat, Env, Pol, Gag with adjuvants (Montanide, Alum and CFA) has potentials as a candidate vaccine against the HIV-1 virus. Vac Res, 2014, 1 (1): 10-15

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

cloning, expression and purification of a novel multi-epitopic hiv-1 vaccine candidate: a preliminary study on immunoreactivity

introduction : designing an effective vaccine against human immunodeficiency virus (hiv)-1 is a global health priority . multi-epitope vaccines offer several potential advantages that may be promising in case of mutable divergent pathogens such as hiv-1. herein, a multiepitopic recombinant protein containing various hiv-1 antigens was expressed in e. coli cells and its immunogenicity in combina...

full text

effects of first language on second language writing-a preliminary contrastive rhetoric study of farsi and english

to explore the idea the investingation proposed, aimed at finding whether the performances of the population of iranians students studying english in an efl context are consistent in l1 and l2 writing taks and whether there is a cross-linguistic transfer in this respect. in this regard the subjects were instructed to write four compositions-two in english and two in farsi-which consisted of an ...

15 صفحه اول

wuthering heights and the concept of marality/a sociological study of the novel

to discuss my point, i have collected quite a number of articles, anthologies, and books about "wuthering heights" applying various ideas and theories to this fantastic story. hence, i have come to believe that gadamer and jauss are rightful when they claim that "the individaul human mind is the center and origin of all meaning," 3 that reading literature is a reader-oriented activity, that it ...

15 صفحه اول

Cloning, Expression, and Refolding of PPE17 Protein of Mycobacterium Tuberculosis as a Promising Vaccine Candidate

Background: Tuberculosis as a global health problem requires special attention in the contexts of prevention and control. Subunit vaccines are promising strategies to protect burdens of tuberculosis via increasing the BCG protection. The present study aimed to design a vaccine study by means of high-throughput expression and correct refolding of recombinant protein PPE17. Methods: We aimed to c...

full text

a study on thermodynamic models for simulation of 1,3 butadiene purification columns

attempts have been made to study the thermodynamic behavior of 1,3 butadiene purification columns with the aim of retrofitting those columns to more energy efficient separation schemes. 1,3 butadiene is purified in two columns in series through being separated from methyl acetylene and 1,2 butadiene in the first and second column respectively. comparisons have been made among different therm...

Cloning and expression of hepatitis E virus ORF2 as a vaccine candidate

Introduction: Hepatitis E virus (HEV) is a fecal-oral transmitting virus which causes a chronic liver disease. ORF2 is an immunogen capsid protein of HEV that has been proposed to be used for Hepatitis E vaccine design. It is a 660-amino acid protein which includes an immunogenic region (residues 112-607). This protein has been expressed in complete and truncated forms, using different expressi...

full text

My Resources

Save resource for easier access later

Save to my library Already added to my library

{@ msg_add @}


Journal title

volume 1  issue 1

pages  10- 15

publication date 2014-08

By following a journal you will be notified via email when a new issue of this journal is published.

Keywords

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023