title: Production of a Locus- and Allele-Specific Monoclonal Antibody for the Characterization of SLA-1*0401 mRNA and Protein Expression Levels in MHC-Defined Microminipigs.
abstract: The class I major histocompatibility complex (MHC) presents self-developed peptides to specific T cells to induce cytotoxity against infection. The MHC proteins are encoded by multiple loci that express numerous alleles to preserve the variability of the antigen-presenting ability in each species. The mechanism regulating MHC mRNA and protein expression at each locus is difficult to analyze because of the structural and sequence similarities between alleles. In this study, we examined the correlation between the mRNA and surface protein expression of swine leukocyte antigen (SLA)-1*0401 after the stimulation of peripheral blood mononuclear cells (PBMCs) by Staphylococcus aureus superantigen toxic shock syndrome toxin-1 (TSST-1). We prepared a monoclonal antibody (mAb) against a domain composed of Y102, L103 and L109 in the α2 domain. The Hp-16.0 haplotype swine possess only SLA-1*0401, which has the mAb epitope, while other haplotypes possess 0 to 3 SLA classical class I loci with the mAb epitopes. When PBMCs from SLA-1*0401 homozygous pigs were stimulated, the SLA-1*0401 mRNA expression level increased until 24 hrs and decreased at 48 hrs. The kinetics of the interferon regulatory transcription factor-1 (IRF-1) mRNA level were similar to those of the SLA-1*0401 mRNA. However, the surface protein expression level continued to increase until 72 hrs. Similar results were observed in the Hp-10.0 pigs with three mAb epitopes. These results suggest that TSST-1 stimulation induced both mRNA and surface protein expression of class I SLA in the swine PBMCs differentially and that the surface protein level was sustained independently of mRNA regulation.
authors.affiliation: Department of Molecular Life Science, Division of Basic Medical Science, Tokai University School of Medicine, Isehara, Kanagawa, Japan.|Institute of Advanced Biosciences, Tokai University, Kanagawa, Japan.
authors.forename: Yoshie
authors.initials: Y
authors.lastname: Kametani
authors.affiliation: Department of Molecular Life Science, Division of Basic Medical Science, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
authors.forename: Shino
authors.initials: S
authors.lastname: Ohshima
authors.affiliation: Department of Molecular Life Science, Division of Basic Medical Science, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
authors.forename: Asuka
authors.initials: A
authors.lastname: Miyamoto
authors.affiliation: Department of Molecular Life Science, Division of Basic Medical Science, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
authors.forename: Atsuko
authors.initials: A
authors.lastname: Shigenari
authors.affiliation: Department of Veterinary Medicine, Faculty of Applied Biological Sciences, Gifu University, Gifu, Gifu, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Takasu
authors.affiliation: Department of Veterinary Medicine, Faculty of Applied Biological Sciences, Gifu University, Gifu, Gifu, Japan.
authors.forename: Noriaki
authors.initials: N
authors.lastname: Imaeda
authors.affiliation: Department of Veterinary Medicine, Faculty of Applied Biological Sciences, Gifu University, Gifu, Gifu, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Matsubara
authors.affiliation: Department of Molecular Life Science, Division of Basic Medical Science, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
authors.forename: Masafumi
authors.initials: M
authors.lastname: Tanaka
authors.affiliation: Department of Molecular Life Science, Division of Basic Medical Science, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Shiina
authors.affiliation: Teaching and Research Support Center, Tokai University School of Medicine, Isehara, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kamiguchi
authors.affiliation: Department of Rheumatology and Clinical Immunology, Clinical Research Center for Allergy and Rheumatology, Sagamihara National Hospital, National Hospital Organization, Sagamihara, Kanagawa, Japan.
authors.forename: Ryuji
authors.initials: R
authors.lastname: Suzuki
authors.affiliation: Department of Veterinary Medicine, Faculty of Applied Biological Sciences, Gifu University, Gifu, Gifu, Japan.
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Kitagawa
authors.affiliation: Department of Molecular Life Science, Division of Basic Medical Science, Tokai University School of Medicine, Isehara, Kanagawa, Japan.|School of Psychiatry and Clinical Neurosciences, The University of Western Australia, Crawley WA, Australia.
authors.forename: Jerzy K
authors.initials: JK
authors.lastname: Kulski
authors.affiliation: Institute of Glycoscience, Tokai University, Hiratsuka, Kanagawa, Japan.
authors.forename: Noriaki
authors.initials: N
authors.lastname: Hirayama
authors.affiliation: Department of Molecular Life Science, Division of Basic Medical Science, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Inoko
authors.affiliation: Department of Molecular Life Science, Division of Basic Medical Science, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
authors.forename: Asako
authors.initials: A
authors.lastname: Ando
mesh_terms: D000483:Alleles; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000918:Antibody Specificity; D001427:Bacterial Toxins / Q000276:immunology*; D004768:Enterotoxins / Q000276:immunology*; D000939:Epitopes / Q000276:immunology; D005802:Genes, MHC Class II; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000276:immunology; D013211:Staphylococcus aureus / Q000276:immunology / Q000378:metabolism; D018089:Superantigens / Q000276:immunology*; D013552:Swine; D013556:Swine, Miniature
chemical_list: D000911:Antibodies, Monoclonal; D001427:Bacterial Toxins; D004768:Enterotoxins; D000939:Epitopes; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D018089:Superantigens; C030107:enterotoxin F, Staphylococcal; C438668:swine leukocyte antigen