title: Intranasal chitosan-DNA vaccines that protect across influenza virus subtypes.
abstract: An egg-free and broadly effective influenza vaccine that can be produced rapidly, adequately and cost-effectively is needed. In this study, chitosan-associated DNAs prepared at various nitrogen/phosphate charge (N/P) ratios were studied for their physicochemical properties, stability, cytotoxicity, and protein expression ability. The chitosan-DNA complexes (chitoplexes) of the N/P ratio 2 had the required characteristics including optimal size range, positive surface charge, high DNA association efficiency, tolerability to DNase digestion and mammalian cell viability compatibility. The N/P ratio 2-chitoplexes revealed the highest green fluorescent protein and luciferase expressions in the transfected mammalian cell cultures and in the mouse lungs. Mice immunized intranasally with the N/P ratio 2-chitoplex vaccines carrying DNAs coding for conserved proteins of influenza virus, i.e., ion channel protein (M2) and/or nucleoprotein (NP), had both mucosal and systemic humoral as well as cell mediated immune responses to the in vivo expressed antigens which conferred protection in mice against the lethal challenges not only with the homologous influenza virus subtype (H1N1), but also the heterologous subtype (H3N2). The chitoplexes should be considered as influenza vaccine candidates especially during the period of high vaccine demand. They are suitable for developing areas of the world where conventional vaccine production capacity is lacking.
authors.affiliation: Department of Manufacturing Pharmacy, Faculty of Pharmacy, Mahidol University, Bangkok 10400, Thailand; Laboratory of Virology and Vaccinology, Division of Biomedical Research, National Institute of Biomedical Innovation, Osaka 567-0085, Japan.
authors.forename: Chompoonuch
authors.initials: C
authors.lastname: Sawaengsak
authors.affiliation: Laboratory of Virology and Vaccinology, Division of Biomedical Research, National Institute of Biomedical Innovation, Osaka 567-0085, Japan; Division of Clinical Virology, Department of Microbiology and Infectious Diseases, Kobe University Graduate School of Medicine, Kobe 657-0013, Japan.
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Mori
authors.affiliation: National Institute of Biomedical Innovation, Osaka 567-0085, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Yamanishi
authors.affiliation: Graduate Program in Biomedical Science, Faculty of Allied Health Sciences, Thammasat University, Pathumthani 12120, Thailand.
authors.forename: Potjanee
authors.initials: P
authors.lastname: Srimanote
authors.affiliation: Laboratory for Research and Technology Development, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.
authors.forename: Wanpen
authors.initials: W
authors.lastname: Chaicumpa
authors.affiliation: Department of Manufacturing Pharmacy, Faculty of Pharmacy, Mahidol University, Bangkok 10400, Thailand.
authors.forename: Ampol
authors.initials: A
authors.lastname: Mitrevej
authors.affiliation: Department of Manufacturing Pharmacy, Faculty of Pharmacy, Mahidol University, Bangkok 10400, Thailand. Electronic address: nuttanan.sin@mahidol.ac.th.
authors.forename: Nuttanan
authors.initials: N
authors.lastname: Sinchaipanid
mesh_terms: D000281:Administration, Intranasal; D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D048271:Chitosan / Q000008:administration & dosage* / Q000737:chemistry; D004285:Dogs; D005260:Female; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology*; D053122:Influenza A Virus, H3N2 Subtype / Q000276:immunology*; D007252:Influenza Vaccines / Q000008:administration & dosage* / Q000737:chemistry; D061985:Madin Darby Canine Kidney Cells; D008807:Mice, Inbred BALB C; D009976:Orthomyxoviridae Infections / Q000517:prevention & control*; D019444:Vaccines, DNA / Q000008:administration & dosage* / Q000737:chemistry
chemical_list: D007252:Influenza Vaccines; D019444:Vaccines, DNA; D048271:Chitosan
keywords: Chitosan self assembly; DNA vaccine; Heterosubtypic immunity; Influenza; Intranasal vaccination; Particulate gene delivery system