title: Vaccination with recombinant 4 × M2e.HSP70c fusion protein as a universal vaccine candidate enhances both humoral and cell-mediated immune responses and decreases viral shedding against experimental challenge of H9N2 influenza in chickens.
abstract: As cellular immunity is essential for virus clearance, it is commonly accepted that no adequate cellular immunity is achieved by all available inactivated HA-based influenza vaccines. Thus, an improved influenza vaccine to induce both humoral and cell-mediated immune responses is urgently required to control LPAI H9N2 outbreaks in poultry farms. M2e-based vaccines have been suggested and developed as a new generation of universal vaccine candidate against influenza A infection. Our previous study have shown that a prime-boost administration of recombinant 4×M2e.HSP70c (r4M2e/H70c) fusion protein compared to conventional HA-based influenza vaccines provided full protection against lethal dose of influenza A viruses in mice. In the present study, the immunogenicity and protective efficacy of (r4M2e/H70c) was examined in chickens. The data reported herein show that protection against H9N2 viral challenge was significantly increased in chickens by injection of r4M2e/H70c compared with injection of conventional HA-based influenza vaccine adjuvanted with MF59 or recombinant 4×M2e (r4M2e) without HSP70c. Oropharyngeal and cloacal shedding of the virus was detected in all of the r4M2e/H70c vaccinated birds at 2 days after challenge, but the titer was low and decreased rapidly to reach undetectable levels at 7 days after challenge. Moreover, comparison of protective efficacy against LPAI H9N2 in birds intramuscularly immunized with r4M2e/H70c likely represented the ability of the M2e-based vaccine in providing cross-protection against heterosubtypic H9N2 challenge and also allowed the host immune system to induce HA-homosubtype neutralizing antibody against H9N2 challenge. This protective immunity might be attributed to enhanced cell-mediated immunity, which is interpreted as increased lymphocytes proliferation, increased levels of Th1-type (IFN-γ) and Th2-type (IL-4) cytokines production and increased CD4(+) to CD8(+) ratios, resulting from the injection of four tandem repeats of the ectodomain of the conserved influenza matrix protein M2 (4×M2e) genetically fused to C-terminus of Mycobacterium tuberculosis HSP70 (mHSP70c).
authors.affiliation: Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, PO Box 14155-3651, Tehran, Iran; Department of Pathobiology, University of Tehran, Faculty of Veterinary Medicine, PO Box 14155-6453, Tehran, Iran.
authors.forename: Mehran
authors.initials: M
authors.lastname: Dabaghian
authors.affiliation: Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, PO Box 14155-3651, Tehran, Iran.
authors.forename: Ali Mohammad
authors.initials: AM
authors.lastname: Latify
authors.affiliation: Department of Biotechnology, Razi Vaccine and Serum Research Institute (RVSRI), PO Box 31975/148, Karaj, Tehran, Iran.
authors.forename: Majid
authors.initials: M
authors.lastname: Tebianian
authors.affiliation: Department of Avian Research, School of Veterinary Medicine, Shiraz University, PO Box 1731, Shiraz, Iran.
authors.forename: Hassan
authors.initials: H
authors.lastname: Nili
authors.affiliation: Department of Medical Virology, Iran University of Medical Sciences, Tehran, Iran.
authors.forename: Ali Reza Tevangar
authors.initials: AR
authors.lastname: Ranjbar
authors.affiliation: Department of Biotechnology, Razi Vaccine and Serum Research Institute (RVSRI), PO Box 31975/148, Karaj, Tehran, Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Mirjalili
authors.affiliation: Department of Biotechnology, Razi Vaccine and Serum Research Institute (RVSRI), PO Box 31975/148, Karaj, Tehran, Iran.
authors.forename: Mashallah
authors.initials: M
authors.lastname: Mohammadi
authors.affiliation: Department of Medical Immunology, Tarbiyat Modares University, Tehran, Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Banihashemi
authors.affiliation: Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, PO Box 14155-3651, Tehran, Iran. Electronic address: sme3422@gmail.com.
authors.forename: Seyyed Mahmoud
authors.initials: SM
authors.lastname: Ebrahimi
mesh_terms: D000818:Animals; D002645:Chickens; D056738:Cross Protection / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005434:Flow Cytometry / Q000662:veterinary; D018840:HSP70 Heat-Shock Proteins / Q000008:administration & dosage / Q000276:immunology*; D006386:Hemagglutination Tests / Q000662:veterinary; D007111:Immunity, Cellular / Q000187:drug effects / Q000276:immunology; D056724:Immunity, Humoral / Q000187:drug effects / Q000276:immunology; D016130:Immunophenotyping / Q000662:veterinary; D053127:Influenza A Virus, H9N2 Subtype / Q000276:immunology*; D007252:Influenza Vaccines / Q000008:administration & dosage / Q000276:immunology*; D005585:Influenza in Birds / Q000517:prevention & control*; D007273:Injections, Intramuscular / Q000662:veterinary; D015847:Interleukin-4 / Q000276:immunology; D011201:Poultry Diseases / Q000517:prevention & control* / Q000821:virology; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000276:immunology; D014611:Vaccination / Q000662:veterinary; D015164:Vaccines, Inactivated / Q000008:administration & dosage / Q000276:immunology; D017201:Virus Shedding / Q000187:drug effects / Q000276:immunology
chemical_list: D018840:HSP70 Heat-Shock Proteins; D007252:Influenza Vaccines; D011993:Recombinant Fusion Proteins; D015164:Vaccines, Inactivated; D015847:Interleukin-4
keywords: Chicken; H9N2; HSP70c; Influenza A universal vaccine; M2e