All of these expressed high-avidity T cell receptors for M1p/HLA*02:01 tetramers, and 2 of the 3 receptors were sequenced. Microrafts with highly active CD8 + T cells were individually transferred to wells of a 96-well plate, using a needle-release device coupled to the microscope. ![]() The rates of target cell death among the individual CD8 + T cells varied greatly however, individual T cells maintained their rates of cytotoxicity throughout the time course of the experiment enabling rapid identification of highly cytotoxic CD8 + T cells. Target cell killing, measured by fluorescence microscopy, was quantified in each microraft. Individual microrafts on a 70 × 70 array were loaded with on average 1 CD8 + cell from the culture and a population of M1p presenting target cells. We have developed a microraft array methodology that automatically measures the ability of individual T cells to kill a population of target cells and viably sorts specific cells into a 96-well plate for expansion.Ī human T cell culture was generated against the influenza M1p antigen. ![]() ![]() The simultaneous measurement of T cell function with recovery of individual T cells would greatly facilitate characterizing antigen-specific responses both in vivo and in model systems.
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