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Intrathymic administration of antigen is a versatile strategy for the induction of tolerance to a wide range of allo-, xeno-, and autoantigens. Interest has now turned to developing methods allowing efficient delivery of antigen to the thymus, and to elucidating the mechanisms responsible for tolerance induction using this route of antigen delivery. Novel approaches for introducing antigen into the thymus, including the use of gene transfer and bone marrow chimeras, have been developed. Recent data utilizing T-cell receptor transgenic animals have shed new light on the mechanisms responsible for acquired intrathymic tolerance.