title: Characterization and immunogenicity in mice of recombinant influenza haemagglutinins produced in Leishmania tarentolae.
abstract: The membrane displayed antigen haemagglutinin (HA) from several influenza strains were expressed in the Leishmania tarentolae system. This non-conventional expression system based on a parasite of lizards, can be readily propagated to high cell density (>10(8)cells/mL) in a simple incubator at 26°C. The genes encoding HA proteins were cloned from six influenza strains, among these being a 2009 A/H1N1 pandemic strain from swine origin, namely A/California/07/09(H1N1). Soluble HA proteins were secreted into the cell culture medium and were easily and successfully purified via a His-Tag domain fused to the proteins. The overall process could be conducted in less than 3 months and resulted in a yield of approximately 1.5-5mg of HA per liter of biofermenter culture after purification. The recombinant HA proteins expressed by L. tarentolae were characterized by dynamic light scattering and were observed to be mostly monomeric. The L. tarentolae recombinant HA proteins were immunogenic in mice at a dose of 10μg when administered twice with an oil-in-water emulsion-based adjuvant. These results suggest that the L. tarentolae expression system may be an alternative to the current egg-based vaccine production.
authors.affiliation: Department of Research and Development, Sanofi Pasteur, 1541 Avenue Marcel Mérieux, 69280 Marcy L'Etoile, France. Electronic address: corinne.pion@sanofipasteur.com.
authors.forename: Corinne
authors.initials: C
authors.lastname: Pion
authors.affiliation: Department of Research and Development, Sanofi Pasteur, 1541 Avenue Marcel Mérieux, 69280 Marcy L'Etoile, France. Electronic address: virginie.courtois@sanofipasteur.com.
authors.forename: Virginie
authors.initials: V
authors.lastname: Courtois
authors.affiliation: Department of Research and Development, Sanofi Pasteur, 1541 Avenue Marcel Mérieux, 69280 Marcy L'Etoile, France. Electronic address: stephanie.husson@sanofipasteur.com.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Husson
authors.affiliation: Department of Research and Development, Sanofi Pasteur, 1541 Avenue Marcel Mérieux, 69280 Marcy L'Etoile, France. Electronic address: marie-clotilde.bernard@sanofipasteur.com.
authors.forename: Marie-Clotilde
authors.initials: MC
authors.lastname: Bernard
authors.affiliation: Department of Research and Development, Sanofi Pasteur, 1541 Avenue Marcel Mérieux, 69280 Marcy L'Etoile, France. Electronic address: marie-claire.nicolai@sanofipasteur.com.
authors.forename: Marie-Claire
authors.initials: MC
authors.lastname: Nicolai
authors.affiliation: Department of Research and Development, Sanofi Pasteur, 1541 Avenue Marcel Mérieux, 69280 Marcy L'Etoile, France. Electronic address: philippe.talaga@sanofipasteur.com.
authors.forename: Philippe
authors.initials: P
authors.lastname: Talaga
authors.affiliation: Department of Research and Development, Sanofi Pasteur, 1541 Avenue Marcel Mérieux, 69280 Marcy L'Etoile, France. Electronic address: emanuelle.trannoy@sanofipasteur.com.
authors.forename: Emanuelle
authors.initials: E
authors.lastname: Trannoy
authors.affiliation: Department of Research and Development, Sanofi Pasteur, 1541 Avenue Marcel Mérieux, 69280 Marcy L'Etoile, France. Electronic address: catherine.moste@sanofipasteur.com.
authors.forename: Catherine
authors.initials: C
authors.lastname: Moste
authors.affiliation: Department of Research and Development, Sanofi Pasteur, 1541 Avenue Marcel Mérieux, 69280 Marcy L'Etoile, France; Technology Research Institute Bioaster, 317 Avenue Jean-Jaurès, 69007 Lyon, France. Electronic address: regis.sodoyer@sanofipasteur.com.
authors.forename: Régis
authors.initials: R
authors.lastname: Sodoyer
authors.affiliation: Department of Research and Development, Sanofi Pasteur, 1541 Avenue Marcel Mérieux, 69280 Marcy L'Etoile, France. Electronic address: isabelle.legastelois@sanofipasteur.com.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Legastelois
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D003001:Cloning, Molecular; D005260:Female; D006385:Hemagglutination Inhibition Tests; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000096:biosynthesis / Q000276:immunology*; D007074:Immunoglobulin G / Q000097:blood; D053118:Influenza A Virus, H1N1 Subtype; D007252:Influenza Vaccines / Q000276:immunology*; D007891:Leishmania / Q000378:metabolism*; D008807:Mice, Inbred BALB C; D009976:Orthomyxoviridae Infections / Q000517:prevention & control*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007074:Immunoglobulin G; D007252:Influenza Vaccines; D011994:Recombinant Proteins
keywords: Haemagglutinin; Influenza; Leishmania tarentolae