Determining specific T cells, present at low frequency, useful, e.g. for diagnosis and development of tumor vaccine, based on reaction with HLA Class I tetramers

摘要

Method for determining specific T cells (A), present at very low frequency. HLA (human leukocyte antigen) Class I tetramers (Te) for a selected T-cell antigen (X) are prepared using avidin or an avidin homolog (or their conjugates), and then used to label a sample, i.e. blood mononuclear cells or an enriched T cell population. The treated sample is pipetted into a 96-well plate that has, as its base, a nitrocellulose membrane coated with antibody (Ab) against avidin or homolog. T cells are (i) released from Te, either mechanically or by addition of antibody or buffer then (ii) rinsed from the plate. Optionally, if no enzyme-coupled avidin (or homolog) has been used, then an enzyme-coupled antibody against avidin (or against a substance coupled to it) is added and the Te left behind are visualized by adding a substrate.