T cell receptor-transgenic primary T cells as a tool for discovery of leukaemia-associated antigens.
Identification of a broad array of leukaemia-associated antigens is a crucial step towards immunotherapy of haematological malignancies. However, it is frequently hampered by the decrease of proliferative potential and functional activity of T cell clones used for screening procedures. Transfer of the genes encoding the T cell receptor (TCR) alpha and beta chains of leukaemia-specific clones into primary T cells may help to circumvent this obstacle. In this study, transfer of two minor histocompatibility antigen (minor H antigen)-specific TCRs was performed and the feasibility of the use of TCR-transgenic T cells for identification of minor H antigens through cDNA library screening was investigated. We found that TCR-transgenic cells acquired the specificity of the original clones and matched their sensitivity. Moreover, the higher scale of cytokine-production by TCR-transgenic T cells permits the detection of either small amounts of antigen-positive cells or cells expressing low amounts of an antigen. When applied in equal numbers, TCR-transgenic T cells and the original T cell clones produced similar results in the screening of a cDNA library. However, the use of increased numbers of TCR-transgenic T cells allowed detection of minute amounts of antigen, barely discernible by the T cell clone. In conclusion, TCR-transfer generates a large amount of functional antigen-specific cells suitable for screening of cDNA expression libraries for identification of cognate antigens.
Ivanov
R
R
Jordan Laboratory for Hemato-Oncology, Department of Hematology, University Medical Center Utrecht, Utrecht, The Netherlands.
Hol
S
S
Aarts
T I
TI
Hagenbeek
A
A
Ebeling
S B
SB
D003001:Cloning, Molecular; D016207:Cytokines / Q000276:immunology; D020869:Gene Expression Profiling; D005822:Genetic Vectors / Q000008:administration & dosage; D006680:HLA Antigens / Q000032:analysis*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000276:immunology; D012190:Retroviridae / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology*; D014161:Transduction, Genetic / Q000379:methods; D019076:Transgenes
D016207:Cytokines; D006680:HLA Antigens; D011948:Receptors, Antigen, T-Cell