title: Humoral and cellular immune responses in sheep immunized with a 22 kilodalton exported protein of Mycobacterium avium subspecies paratuberculosis.
abstract: An immunogenic 22 kilodalton exported Mycobacterium avium subspecies paratuberculosis (MAP) lipoprotein (P22) was previously identified, and found to belong to the LppX/LprAFG family of mycobacterial lipoproteins. N-terminal polyhistidine-tagged P22 was produced and purified from Escherichia coli. Antibody recognition of P22, and interferon-gamma (IFN-gamma) responses in vitro using blood from a sheep vaccinated with Neoparasec, confirmed its immunogenicity. To evaluate the immunogenicity of P22 in vivo, five sheep were immunized with a single dose containing 0.8 mg recombinant P22 protein in adjuvant. Blood was collected at 4, 13 and 29 weeks post-immunization (p.i.) and tested for anti-P22 antibodies and P22-specific IFN-gamma production. P22-specific antibodies were detected by Western blot analysis in all five Neoparasec-immunized sheep at the three time points. Three out of five P22-immunized sheep produced P22-specific antibodies for up to 13 weeks p.i., and two gave a response at 29 weeks p.i. Recombinant P22 was able to stimulate significant IFN-gamma production in blood of P22-immunized sheep at 13 and 29 weeks p.i. Recombinant P22 also elicited an IFN-gamma response in blood of sheep immunized with Neoparasec.
authors.affiliation: Institute of Veterinary, Animal and Biomedical Science, Massey University, Private Bag 11 222, Palmerston North, New Zealand.
authors.forename: Rachael C
authors.initials: RC
authors.lastname: Rigden
authors.affiliation: Institute of Veterinary, Animal and Biomedical Science, Massey University, Private Bag 11 222, Palmerston North, New Zealand.
authors.forename: Dakshina M
authors.initials: DM
authors.lastname: Jandhyala
authors.affiliation: Institute of Veterinary, Animal and Biomedical Science, Massey University, Private Bag 11 222, Palmerston North, New Zealand.
authors.forename: Chris
authors.initials: C
authors.lastname: Dupont
authors.affiliation: Institute of Veterinary, Animal and Biomedical Science, Massey University, Private Bag 11 222, Palmerston North, New Zealand.
authors.forename: Dianna
authors.initials: D
authors.lastname: Crosbie-Caird
authors.affiliation: Institute of Veterinary, Animal and Biomedical Science, Massey University, Private Bag 11 222, Palmerston North, New Zealand.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Lopez-Villalobos
authors.affiliation: Unité de Génétique Mycobactérienne, Institut Pasteur, 25 rue du Dr Roux, 75724 Paris Cedex 15, France.
authors.forename: Norihiro
authors.initials: N
authors.lastname: Maeda
authors.affiliation: Unité de Génétique Mycobactérienne, Institut Pasteur, 25 rue du Dr Roux, 75724 Paris Cedex 15, France.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Gicquel
authors.affiliation: Unité de Génétique Mycobactérienne, Institut Pasteur, 25 rue du Dr Roux, 75724 Paris Cedex 15, France.|Institute of Veterinary, Animal and Biomedical Science, Massey University, Private Bag 11 222, Palmerston North, New Zealand.
authors.forename: Alan
authors.initials: A
authors.lastname: Murray
mesh_terms: D000276:Adjuvants, Immunologic; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood* / Q000276:immunology; D000918:Antibody Specificity; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*; D001428:Bacterial Vaccines / Q000008:administration & dosage / Q000276:immunology*; D015153:Blotting, Western; D004926:Escherichia coli / Q000378:metabolism; D007114:Immunization; D007279:Injections, Subcutaneous; D007371:Interferon-gamma / Q000097:blood* / Q000276:immunology; D008074:Lipoproteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*; D008297:Male; D008970:Molecular Weight; D009162:Mycobacterium avium / Q000737:chemistry / Q000276:immunology*; D009821:Oils; D011994:Recombinant Proteins / Q000276:immunology; D012756:Sheep; D014376:Tuberculosis / Q000097:blood / Q000276:immunology*; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000276:immunology; D014867:Water
chemical_list: D000276:Adjuvants, Immunologic; D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D008074:Lipoproteins; D009821:Oils; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D014867:Water; D007371:Interferon-gamma