The study of monoclonal human T-cell populations has had a fundamental impact on our current knowledge of the function, specificity, and mechanisms of activation of these cells. The frequency of antigen-specific T cells in peripheral blood is low, necessitating several enrichment steps prior to the isolation of individual clones. Two different methods of limiting dilution cloning are described in this chapter, the choice of which depends on the availability of starting materials. After isolation, T-cell clones are expanded and then tested for specificity and cryopreserved, both of which are described.Recently, time-saving variations to the above methods have emerged. However, they require the use of sophisticated equipment and reagents, making them less economical than the established techniques.
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