Telomeres are unique DNA-protein complexes at the terminals of chromosomes that appear to play a critical role in protecting chromosomal integrity and in maintaining cellular replicative potential. Telomerase is a ribonuclear protein that is capable of elongating telomeres by the addition of telomeric hexanucleotide repeats and therefore contributing to the capacity for cell replication. Telomerase activity is expressed in human germline cells and malignant cells, and it has recently been demonstrated that telomerase activity is highly regulated in normal lymphocytes at specific stages of development and activation. However, these studies have not elucidated whether telomerase activity is regulated at the level of specific gene expression or whether the regulation of telomerase RNA template (hTR) and/or protein components contributes to the regulation of telomerase activity in normal somatic cells. To characterize at a molecular level the regulation of telomerase expression in human T lymphocytes, we analyzed the expression of hTR during lineage development and after in vitro activation. It was found that hTR is expressed in subsets of thymocytes with strong telomerase activity at levels that are consistently higher (1.5 times; p < 0.01) than those found in peripheral blood resting T cells. In addition, hTR is up-regulated two- to fivefold in peripheral blood naive and memory CD4+ T cells after in vitro activation with anti-CD3 plus anti-CD28. These results establish that hTR expression is regulated in normal human T cells during lineage development and after activation, and indicate that regulation of hTR expression may contribute to the regulation of telomerase activity in normal lymphoid cells.