Production of a Locus- and Allele-Specific Monoclonal Antibody for the Characterization of SLA-10401 mRNA and Protein Expression Levels in MHC-Defined Microminipigs

• Published in: PloS one Vol. 11; no. 10; p. e0164995
• Main Authors: Kametani, Yoshie, Ohshima, Shino, Miyamoto, Asuka, Shigenari, Atsuko, Takasu, Masaki, Imaeda, Noriaki, Matsubara, Tatsuya, Tanaka, Masafumi, Shiina, Takashi, Kamiguchi, Hiroshi, Suzuki, Ryuji, Kitagawa, Hitoshi, Kulski, Jerzy K, Hirayama, Noriaki, Inoko, Hidetoshi, Ando, Asako
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 19.10.2016; Public Library of Science (PLoS)
• Subjects: Alleles; Animals; Antibodies, Monoclonal - metabolism; Antibody Specificity; Antigenic determinants; Antigens; Bacterial Toxins - immunology; Biology and Life Sciences; Coding; Correlation analysis; Cytokines; Enterotoxins - immunology; Epitopes; Epitopes - immunology; Gene expression; Genes, MHC Class II; Haplotypes; Histocompatibility Antigens Class II - genetics; Histocompatibility Antigens Class II - immunology; Histocompatibility Antigens Class II - metabolism; Hogs; Immunoglobulins; Immunologic factors; Interferon; Interferon regulatory factor 1; Kinetics; Leukocytes; Leukocytes (mononuclear); Leukocytes, Mononuclear - immunology; Life sciences; Livestock; Loci; Lymphocytes; Lymphocytes T; Major histocompatibility complex; Medicine and Health Sciences; Messenger RNA; MHC antibodies; Monoclonal antibodies; Peptides; Peripheral blood mononuclear cells; Proteins; Research and Analysis Methods; Rheumatology; Septic shock; Staphylococcal enterotoxin F; Staphylococcus aureus; Staphylococcus aureus - immunology; Staphylococcus aureus - metabolism; Staphylococcus infections; Stimulation; Superantigens - immunology; Swine; Swine, Miniature; T cell receptors; T cells; Toxic shock syndrome; Veterinary medicine; Japan
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0164995

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.