Journal of Isfahan Medical School

Journal of Isfahan Medical School

Prokaryotic Production of Influenza Virus M2 Protein Fused to Leishmania Major HSP70 in Order to Prepare an Effective Flu Vaccine

Document Type : Original Article (s)

Authors
Siavash Chalabiani 1
Fatemeh Fotouhi 2
Amir Ghaemi 3
Maryam Saleh 4
Behrokh Farahmand 2
Mohammad Ali Alavi-Esfahani 1
Mansoureh Tabatabaian 5
Ali Torabi 5
1 Department of Microbiology, School of Biological Science, Islamic Azad University, Qom Branch, Qom, Iran
2 Assistant Professor, Department of Virology, Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran
3 Assistant Professor, Department of Medical Virology and Immunology, School of Medicine, Golestan University of Medical Sciences, Gorgan, Iran
4 Department of Virology, Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran
5 Lab Instructor, Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran
Abstract
Background: Influenza is a major cause of morbidity and mortality in the world. Permanent antigenic variation of influenza virus A causes a major concern to develop influenza vaccine. Now, some researchers are focusing on conserved antigens. The M2 protein is a proton-selective ion channel, integral in the viral envelope of the influenza virus A and allows the virus to enter and cause an infection in the host cells. This protein is conserved among all types of influenza virus A and an appropriate target for the development of influenza vaccine with broad-spectrum protection. In this study, Leishmania major heat shock protein-70 fused to M2 protein to prepare an effective vaccine against influenza virus A.Methods: Lm. HSP70 gene was cloned into BamHI and HindIII sites of pET28a vector. Influenza virus M2 gene was amplified via polymerase chain reaction (PCR) using specific primers and cloned into dephosphorylated linear pET28a vector upstream of Leishmania major HSP70 gene. The confirmed construct was transformed into Escherichia coli BL21 and protein expression was induced using isopropyl β-D-1-thiogalactopyranoside (IPTG).Findings: The recombinant plasmids were confirmed via colony PCR, restriction enzyme analysis and sequencing. The result of sequencing revealed that the M2 gene was properly cloned into pET28a-HSP70 in the right frame to 6xhis tag. The protein expression was determined using Western blot analysis.Conclusion: Binding of HSP to the desired antigen induces increased level of immune responses. Hence, the chimer protein prepared in this study, could be an appropriate vaccine candidate to prevent influenza infection. The immunogenicity of this chimer protein with different formulation is going to evaluate in animal models. To investigate the effect of HSP on M2 immunogenicity, this chimer protein will be evaluated in future projects.
Keywords
Influenza virus A
Chimer protein
M2
HSP70
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Journal of Isfahan Medical School
Volume 32, Issue 288 - Serial Number 288
May and June 2014
Pages 841-853

  • Receive Date 06 May 2014
  • Accept Date 06 April 2022