title: HLA class I mono-specific APCs and target cells: a method to standardise in vitro CD8+ T cell expansion and functional assays.
abstract: The introduction of in vitro T cell expansion and assay methods that are robust and easy to use would be welcome in cancer vaccine and infectious disease research. By coating HLA class I--ve B cells with recombinant HLA class I peptide complexes, we are able to produce antigen presenting cells and target cells expressing a single defined antigen in the context of costimulatory and adhesion molecules. HLA class I mono-specific cells promoted the in vitro expansion of CMV epitope specific CD8+ T cells from 0.03% to 30.6% in 2 weeks, which was comparable to using peptide-loaded dendritic cells. The HLA class I mono-specific cells were also shown to promote in vitro antigen specific T cell function in assays based on measuring cytokine production and cytolytic activity. HLA class I mono-specific cells are simple to prepare, can be used with any recombinant HLA class I allele/peptide combination and should provide a useful system for in vitro T cell expansion and functional analysis.
authors.affiliation: Anthony Nolan Research Institute, Royal Free Hospital, Pond Street, Hampstead, London NW3 2QG, United Kingdom.
authors.forename: Linda D
authors.initials: LD
authors.lastname: Barber
authors.forename: Susan
authors.initials: S
authors.lastname: Jordan
authors.forename: Alison M E
authors.initials: AM
authors.lastname: Whitelegg
authors.forename: J Alejandro
authors.initials: JA
authors.lastname: Madrigal
authors.forename: Philip
authors.initials: P
authors.lastname: Savage
mesh_terms: D000938:Antigen-Presenting Cells / Q000502:physiology*; D001402:B-Lymphocytes / Q000502:physiology*; D018414:CD8-Positive T-Lymphocytes / Q000502:physiology*; D006680:HLA Antigens / Q000276:immunology / Q000502:physiology*; D015395:Histocompatibility Antigens Class I; D006801:Humans; D046509:Tissue Culture Techniques / Q000379:methods*
chemical_list: D006680:HLA Antigens; D015395:Histocompatibility Antigens Class I