title: Adsorption of recombinant poxvirus L1-protein to aluminum hydroxide/CpG vaccine adjuvants enhances immune responses and protection of mice from vaccinia virus challenge.
abstract: The stockpiling of live vaccinia virus vaccines has enhanced biopreparedness against the intentional or accidental release of smallpox. Ongoing research on future generation smallpox vaccines is providing key insights into protective immune responses as well as important information about subunit-vaccine design strategies. For protein-based recombinant subunit vaccines, the formulation and stability of candidate antigens with different adjuvants are important factors to consider for vaccine design. In this work, a non-tagged secreted L1-protein, a target antigen on mature virus, was expressed using recombinant baculovirus technology and purified. To identify optimal formulation conditions for L1, a series of biophysical studies was performed over a range of pH and temperature conditions. The overall physical stability profile was summarized in an empirical phase diagram. Another critical question to address for development of an adjuvanted vaccine was if immunogenicity and protection could be affected by the interactions and binding of L1 to aluminum salts (Alhydrogel) with and without a second adjuvant, CpG. We thus designed a series of vaccine formulations with different binding interactions between the L1 and the two adjuvants, and then performed a series of vaccination-challenge experiments in mice including measurement of antibody responses and post-challenge weight loss and survival. We found that better humoral responses and protection were conferred with vaccine formulations when the L1-protein was adsorbed to Alhydrogel. These data demonstrate that designing vaccine formulation conditions to maximize antigen-adjuvant interactions is a key factor in smallpox subunit-vaccine immunogenicity and protection.
authors.affiliation: Perelman School of Medicine at the University of Pennsylvania, Department of Medicine, Division of Infectious Diseases, Philadelphia, PA 19104-6073, United States.
authors.forename: Yuhong
authors.initials: Y
authors.lastname: Xiao
authors.forename: Yuhong
authors.initials: Y
authors.lastname: Zeng
authors.forename: Edward
authors.initials: E
authors.lastname: Alexander
authors.forename: Shyam
authors.initials: S
authors.lastname: Mehta
authors.forename: Sangeeta B
authors.initials: SB
authors.lastname: Joshi
authors.forename: George W
authors.initials: GW
authors.lastname: Buchman
authors.forename: David B
authors.initials: DB
authors.lastname: Volkin
authors.forename: C Russell
authors.initials: CR
authors.lastname: Middaugh
authors.forename: Stuart N
authors.initials: SN
authors.lastname: Isaacs
mesh_terms: D000276:Adjuvants, Immunologic / Q000737:chemistry / Q000494:pharmacology; D000327:Adsorption; D000536:Aluminum Hydroxide / Q000737:chemistry / Q000276:immunology*; D000818:Animals; D000917:Antibody Formation; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000276:immunology*; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D011212:Poxviridae / Q000276:immunology; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology; D012900:Smallpox Vaccine / Q000737:chemistry / Q000276:immunology; D022223:Vaccines, Subunit / Q000276:immunology; D014615:Vaccinia / Q000276:immunology / Q000517:prevention & control*; D014616:Vaccinia virus / Q000276:immunology*; D014758:Viral Core Proteins / Q000737:chemistry / Q000276:immunology*
chemical_list: D000276:Adjuvants, Immunologic; D004268:DNA-Binding Proteins; C109350:I1 protein, vaccinia virus; D011994:Recombinant Proteins; D012900:Smallpox Vaccine; D022223:Vaccines, Subunit; D014758:Viral Core Proteins; D000536:Aluminum Hydroxide