Use of Epstein-Barr virus-based vectors for expression of thyroid auto-antigens in human B-lymphoblastoid cell lines.
The TSH-receptor (TSH-R) and thyroid peroxidase (TPO) are targets of autoantibody production in the autoimmune thyroid disease, Graves' disease, and are also likely to be the target of T-cell responses. To facilitate the analysis of T-cell responses we have investigated a system that allows expression of these autoantigens as recombinant proteins in autologous cells. Human B-lymphoblastoid cell lines (B-LCL), which are known to present antigen to autologous T cells, were transfected with constructs directing the expression of human TSH-R and TPO. The constructs utilized an expression vector replicating under the control of EBV-derived sequences that is maintained episomally in transfected cells. Both proteins were shown to be expressed by transfected B-LCL and present on the cell surface. Such transfected B-LCL could be used for the derivation, screening and characterization of autologous T-cell clones against thyroid autoantigens.
McIntosh
R S
RS
Department of Medicine, Medical School, University of Newcastle upon Tyne, UK.
Mulcahy
A F
AF
Hales
J M
JM
Diamond
A G
AG
D001324:Autoantigens / Q000235:genetics*; D001402:B-Lymphocytes / Q000276:immunology; D001483:Base Sequence; D005822:Genetic Vectors; D006111:Graves Disease / Q000276:immunology; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D007453:Iodide Peroxidase / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D011989:Receptors, Thyrotropin / Q000235:genetics* / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology; D013961:Thyroid Gland / Q000276:immunology; D014162:Transfection; D014407:Tumor Cells, Cultured
D001324:Autoantigens; D015345:Oligonucleotide Probes; D011989:Receptors, Thyrotropin; D011993:Recombinant Fusion Proteins; D007453:Iodide Peroxidase