title: Cell surface expression of MR1B, a splice variant of the MHC class I-related molecule MR1, revealed with antibodies.
abstract: The major histocompatibility complex (MHC) class I-related molecule, MR1, is highly conserved in mammals and can present bacteria-derived vitamin B metabolites to mucosal-associated invariant T (MAIT) cells, possibly having important defense function in the microbial infection. MR1B is a splice variant of MR1 and possesses an intriguing domain structure with only two extracellular domains resembling some NKG2D ligand molecules. Thus far, cell surface expression of MR1B could not be analyzed with flow cytometry due to a lack of appropriate antibodies reactive with MR1B. Here we clarified the expression of MR1B recombinant protein on the cell surface of the transfected cells by flow cytometry analyses using the antiserum against MR1. Consistently, MR1B tagged with FLAG peptide at the N-terminus also could be detected with anti-FLAG monoclonal antibodies. Our result showed that MR1B can be recognized on the cell surface by macromolecules such as antibodies, indicating its potential of interaction with certain receptor(s). We discuss possibility of interaction of MR1B and/or the full-length MR1 with some receptor(s) other than αβ T cell receptor (TCR) of MAIT cells based on the highly conserved characteristic residues of the ligand-binding domains of MR1 and its MAIT cells αβTCR footprints.
authors.affiliation: Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi 470-1192, Japan.
authors.forename: Hisateru
authors.initials: H
authors.lastname: Yamaguchi
authors.affiliation: Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi 470-1192, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Tsukamoto
authors.affiliation: Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi 470-1192, Japan. Electronic address: keihashi@fujita-hu.ac.jp.
authors.forename: Keiichiro
authors.initials: K
authors.lastname: Hashimoto
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D002462:Cell Membrane / Q000276:immunology*; D057809:HEK293 Cells; D015395:Histocompatibility Antigens Class I / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D015778:Minor Histocompatibility Antigens; D020033:Protein Isoforms / Q000235:genetics / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D015395:Histocompatibility Antigens Class I; C094840:MR1 protein, human; D015778:Minor Histocompatibility Antigens; D020033:Protein Isoforms
keywords: CDR; Cell surface expression; HLA; MAIT; MHC; MR1; NK; NKT; Splice variant; T cell receptor; TCR; complementarity determining region; human leukocyte antigen; major histocompatibility complex; mucosal-associated invariant T; natural killer; natural killer T; β(2)-m; β(2)-microglobulin