title: Production and in vivo testing of a recombinant bovine IL-12 as an adjuvant for Salmonella typhimurium vaccination in calves.
abstract: A recombinant bovine interleukin-12 (boIL-12) that contains a histidine hexamer, rboIL-12His, was produced, purified and administered to calves. We first tried the purification of heterodimer IL-12 from a mixture of p40 homodimer, p40 monomer, and p40-p35 heterodimer with a p35 subunit tagged with a histidine hexamar at its C-terminal (p35His). A recombinant baculovirus expressing p35His was generated and used for superinfection with a recombinant baculovirus expressing p40 subunit. The expressed subunits, p40 and p35His, were assembled into a 70kDa heterodimer in insect cells, released into culture medium, and then purified using a nickel chelate column. The purified rboIL-12His was bioactive for induction of IFN-gamma in bovine peripheral blood mononuclear cells (PBMCs) in vitro. The purified rboIL-12His was then administered to calves with inactivated Salmonella Typhimurium (ST). When sera were assayed by ELISA, specific anti-ST IgG1 antibodies were detected in all ST immunized calves, but, specific anti-ST IgG2 antibodies were detected only in calves administered ST along with rboIL-12His, indicating a possible switch to a Th1 response. Administration of commercially available Salmonella vaccine did not elicit IgG2 antibodies in calves. These results suggest that co-administration of IL-12 with inactivated ST cells could induce a Th1-type response in calves.
authors.affiliation: Laboratory of Poultry Diseases, School of Veterinary Medicine and Animal Sciences, Kitasato University, Towada, 034-8628, Aomori, Japan. takehara@vmas.kitasato-u.ac.jp
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Takehara
authors.forename: Reiko
authors.initials: R
authors.lastname: Kikuma
authors.forename: Satoko
authors.initials: S
authors.lastname: Ishikawa
authors.forename: Makiyo
authors.initials: M
authors.lastname: Kamikawa
authors.forename: Tomoshi
authors.initials: T
authors.lastname: Nagata
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Yokomizo
authors.forename: Masayuki
authors.initials: M
authors.lastname: Nakamura
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage* / Q000302:isolation & purification; D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D002417:Cattle / Q000276:immunology*; D002418:Cattle Diseases / Q000276:immunology / Q000517:prevention & control; D019281:Dimerization; D007074:Immunoglobulin G / Q000096:biosynthesis; D007371:Interferon-gamma / Q000096:biosynthesis; D018664:Interleukin-12 / Q000008:administration & dosage* / Q000096:biosynthesis / Q000302:isolation & purification; D008297:Male; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000096:biosynthesis / Q000302:isolation & purification; D012481:Salmonella Infections, Animal / Q000276:immunology / Q000517:prevention & control; D022562:Salmonella Vaccines / Q000008:administration & dosage*; D012486:Salmonella typhimurium / Q000276:immunology*; D018417:Th1 Cells / Q000276:immunology; D015164:Vaccines, Inactivated / Q000008:administration & dosage
chemical_list: D000276:Adjuvants, Immunologic; D000907:Antibodies, Bacterial; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D022562:Salmonella Vaccines; D015164:Vaccines, Inactivated; D018664:Interleukin-12; D007371:Interferon-gamma