Enrichment of terminal deoxynucleotidyl transferase activity by cell separation.
Terminal deoxynucleotidyl transferase is a unique DNA polymerase that can carry out DNA synthesis on an initiator molecule in the absence of a template. The usefulness of this enzyme as a biological marker for following patients during treatment and remission has been suggested. The potential usefulness of this enzyme in predicting the onset of relapse before any morphological indications has been demonstrated in chronic myelogenous leukemia patients in blast phase of the disease. In order to be able to detect low levels of TdT activity especially during remission phase, we have used cell separation techniques which can enrich cell populations containing TdT activity. A number of cell separation techniques have been developed to separate different cell types. We have used the techniques of unit gravity sedimentation and free flow electrophoresis to achieve enrichment of TdT positive cell populations. Our results show that up to 20 fold enrichment of TdT activity in normal human bone marrow can be accomplished by using cell separation techniques. With the use of free flow electrophoresis, we have achieved enrichment of TdT positive cell populations from normal human bone marrow, cells from patients with acute lymphoblastic leukemia and chronic myelogenous leukemia in blast phase of the disease. No TdT positive cells were detected in patients with acute myelogenous leukemia. These cell separation techniques should prove to be useful in early detection of relapse in patients in remission.
D001402:B-Lymphocytes / Q000201:enzymology; D001853:Bone Marrow / Q000201:enzymology / Q000473:pathology; D002460:Cell Line; D002469:Cell Separation / Q000379:methods; D004796:Clinical Enzyme Tests; D004253:DNA Nucleotidylexotransferase / Q000378:metabolism*; D004254:DNA Nucleotidyltransferases / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D004586:Electrophoresis; D006801:Humans; D007945:Leukemia, Lymphoid / Q000175:diagnosis* / Q000201:enzymology / Q000473:pathology; D013601:T-Lymphocytes / Q000201:enzymology
D004254:DNA Nucleotidyltransferases; D004253:DNA Nucleotidylexotransferase; D004259:DNA-Directed DNA Polymerase