title: Comparison of adjuvants for a spray freeze-dried whole inactivated virus influenza vaccine for pulmonary administration.
abstract: Stable vaccines administered to the lungs by inhalation could circumvent many of the problems associated with current immunizations against respiratory infections. We earlier provided proof of concept in mice that pulmonary delivered whole inactivated virus (WIV) influenza vaccine formulated as a stable dry powder effectively elicits influenza-specific antibodies in lung and serum. Yet, mucosal IgA, considered particularly important for protection at the site of virus entry, was poorly induced. Here we investigate the suitability of various Toll-like receptor (TLR) ligands and the saponin-derived compound GPI-0100 to serve as adjuvant for influenza vaccine administered to the lungs as dry powder. The TLR ligands palmitoyl-3-cysteine-serine-lysine-4 (Pam3CSK4), monophosphoryl lipid A (MPLA) and CpG oligodeoxynucleotides (CpG ODN) as well as GPI-0100 tolerated the process of spray freeze-drying well. While Pam3CSK4 had no effect on systemic antibody titers, all the other adjuvants significantly increased influenza-specific serum and lung IgG titers. Yet, only GPI-0100 also enhanced mucosal IgA titers. Moreover, only GPI-0100-adjuvanted WIV provided partial protection against heterologous virus challenge. Pulmonary immunization with GPI-0100-adjuvanted vaccine did not induce an overt inflammatory response since influx of neutrophils and production of inflammatory cytokines were moderate and transient and lung histology was normal. Our results indicate that a GPI-0100-adjuvanted dry powder influenza vaccine is a safe and effective alternative to current parenteral vaccines.
authors.affiliation: Department of Medical Microbiology, University of Groningen and University Medical Center Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands.
authors.forename: Harshad P
authors.initials: HP
authors.lastname: Patil
authors.affiliation: Department of Pharmaceutical Technology and Biopharmacy, University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands.
authors.forename: Senthil
authors.initials: S
authors.lastname: Murugappan
authors.affiliation: Department of Medical Microbiology, University of Groningen and University Medical Center Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands.
authors.forename: Jacqueline
authors.initials: J
authors.lastname: de Vries-Idema
authors.affiliation: Department of Medical Microbiology, University of Groningen and University Medical Center Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands.
authors.forename: Tjarko
authors.initials: T
authors.lastname: Meijerhof
authors.affiliation: Department of Medical Microbiology, University of Groningen and University Medical Center Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands.
authors.forename: Aalzen
authors.initials: A
authors.lastname: de Haan
authors.affiliation: Department of Pharmaceutical Technology and Biopharmacy, University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands.
authors.forename: Henderik W
authors.initials: HW
authors.lastname: Frijlink
authors.affiliation: Department of Medical Microbiology, University of Groningen and University Medical Center Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands.
authors.forename: Jan
authors.initials: J
authors.lastname: Wilschut
authors.affiliation: Department of Pharmaceutical Technology and Biopharmacy, University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands.
authors.forename: Wouter L J
authors.initials: WL
authors.lastname: Hinrichs
authors.affiliation: Department of Medical Microbiology, University of Groningen and University Medical Center Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands. Electronic address: a.l.w.huckriede@umcg.nl.
authors.forename: Anke
authors.initials: A
authors.lastname: Huckriede
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage* / Q000737:chemistry; D000280:Administration, Inhalation; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D015415:Biomarkers / Q000097:blood; D002478:Cells, Cultured; D002626:Chemistry, Pharmaceutical; D018899:CpG Islands; D016207:Cytokines / Q000378:metabolism; D005260:Female; D005612:Freeze Drying; D018836:Inflammation Mediators / Q000378:metabolism; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology*; D007252:Influenza Vaccines / Q000008:administration & dosage* / Q000737:chemistry / Q000276:immunology; D008024:Ligands; D008050:Lipid A / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000737:chemistry / Q000276:immunology; D055666:Lipopeptides / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology; D008807:Mice, Inbred BALB C; D009841:Oligonucleotides / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology; D011208:Powders; D020545:Respiratory Mucosa / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D012503:Saponins / Q000008:administration & dosage* / Q000737:chemistry / Q000276:immunology; D013678:Technology, Pharmaceutical / Q000379:methods; D051193:Toll-Like Receptors / Q000819:agonists / Q000378:metabolism; D014611:Vaccination; D015164:Vaccines, Inactivated / Q000008:administration & dosage
chemical_list: D000276:Adjuvants, Immunologic; D000914:Antibodies, Viral; D015415:Biomarkers; D016207:Cytokines; C484998:GPI0100; D018836:Inflammation Mediators; D007252:Influenza Vaccines; D008024:Ligands; D008050:Lipid A; D055666:Lipopeptides; D009841:Oligonucleotides; C508075:Pam(3)CSK(4) peptide; D011208:Powders; D012503:Saponins; D051193:Toll-Like Receptors; D015164:Vaccines, Inactivated; C048436:monophosphoryl lipid A
keywords: Adjuvants; Pulmonary vaccination; Spray freeze-drying; Whole inactivated virus