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The aims of this project are to determine the phenotype and antileukemic activity of activated bone marrow infiltrating leukemia (MIL) and compare them to activated peripheral blood lymphocytes from patients with chronic myelogenous leukemia (CML) on imatinib or other tyrosine kinase inhibitor therapy. Bone marrow and peripheral blood specimens were obtained from CML patients who had at least a minor cytogenetic response. The phenotype of MILs and peripheral blood lymphocytes (PBL) was analyzed by flow cytometry. MILs and PBLs were expanded and activated with anti CD3/CD28 magnetic beads in culture for 10 days. Activated MILs and PBLs were characterized by flow cytometry and tested for antileukematic activity in a colony suppression assay by coculture with CD34+ bone marrow progenitors at varying CD3:CD34 ratios in methycullulose medium. Analysis of the phenotype of MILs from four patients showed that MILs are predominantly comprised of effector memory T cells. These MILs could be expanded effectively without change in phenotype. MILs as well as activated PBLs showed ability to suppress CML progenitor growth in vitro.