title: Mixed functional characteristics correlating with TCR-ligand koff -rate of MHC-tetramer reactive T cells within the naive T-cell repertoire.
abstract: The low frequency of antigen-specific naïve T cells has challenged numerous laboratories to develop various techniques to study the naïve T-cell repertoire. Here, we combine the generation of naïve repertoire-derived antigen-specific T-cell lines based on MHC-tetramer staining and magnetic-bead enrichment with in-depth functional assessment of the isolated T cells. Cytomegalovirus (CMV) specific T-cell lines were generated from seronegative individuals. Generated T-cell lines consisted of a variety of immunodominant CMV-epitope-specific oligoclonal T-cell populations restricted to various HLA-molecules (HLA-A1, A2, B7, B8, and B40), and the functional and structural avidity of the CMV-specific T cells was studied. Although all CMV-specific T cells were isolated based on their reactivity toward a specific peptide-MHC complex, we observed a large variation in the functional avidity of the MHC-tetramer positive T-cell populations, which correlated with the structural avidity measured by the recently developed Streptamer koff -rate assay. Our data demonstrate that MHC-tetramer staining is not always predictive for specific T-cell reactivity, and challenge the sole use of MHC-tetramers as an indication of the peripheral T-cell repertoire, independent of the analysis of functional activity or structural avidity parameters.
authors.affiliation: Department of Hematology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Pleun
authors.initials: P
authors.lastname: Hombrink
authors.forename: Yotam
authors.initials: Y
authors.lastname: Raz
authors.forename: Michel G D
authors.initials: MG
authors.lastname: Kester
authors.forename: Renate
authors.initials: R
authors.lastname: de Boer
authors.forename: Bianca
authors.initials: B
authors.lastname: Weißbrich
authors.forename: Peter A
authors.initials: PA
authors.lastname: von dem Borne
authors.forename: Dirk H
authors.initials: DH
authors.lastname: Busch
authors.forename: Ton N M
authors.initials: TN
authors.lastname: Schumacher
authors.forename: J H Frederik
authors.initials: JH
authors.lastname: Falkenburg
authors.forename: Mirjam H M
authors.initials: MH
authors.lastname: Heemskerk
mesh_terms: D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D003587:Cytomegalovirus / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D015788:HLA-A1 Antigen / Q000276:immunology; D015789:HLA-A2 Antigen / Q000276:immunology; D059947:HLA-B40 Antigen / Q000276:immunology; D015793:HLA-B7 Antigen / Q000276:immunology; D015795:HLA-B8 Antigen / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D008213:Lymphocyte Activation / Q000276:immunology*; D011948:Receptors, Antigen, T-Cell / Q000276:immunology*; D016176:T-Lymphocyte Subsets / Q000276:immunology
chemical_list: D018984:Epitopes, T-Lymphocyte; D015788:HLA-A1 Antigen; D015789:HLA-A2 Antigen; D059947:HLA-B40 Antigen; D015793:HLA-B7 Antigen; D015795:HLA-B8 Antigen; D011948:Receptors, Antigen, T-Cell; D007371:Interferon-gamma
keywords: CD8 T cells; Immune responses; MHC; Naive cells; TCR