Biochemical Markers for Human Leukemia and Cell Differentiation
Biological markers that can be utilized in following patients during treatment and remission and are differentiation specific have been examined. Two such markers are: (1) terminal deoxynucleotidyl transferase (TdT) and (2) a histone polypeptide (HP). TdT is a unique DNA polymerase that can carry out DNA synthesis on an initiator molecule in the absence of a template. 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. 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 30 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. A differentiation specific histone polypeptide (HP) with an apparent molecular weight of 12,500 has been identified among the acid extractable chromosomal proteins of a human myeloid cell line, HL60, treated with dimethyl sulfoxide (DMSO). The relative increase in the amount of HP in DMSO treated HL60 cells correlated with a concomitant decrease in histone H2A. Fingerprint analysis of tryptic digests shows HP and histone H2A are related, HP is generated from endogenous and exogenous H2A and is probably produced by a proteolytic enzyme associated with the chromatin of the differentiated HL60 cells. HP is not detected in a number of human or mouse hematopoietic tissues and cell lines. However, it is present in leukocytes from patients with acute leukemia. These results indicate that the observed level of HP in HL60 cells is related to the stage of differentiation and that HP is a potential biological markers for human acute leukemia.
KeywordsAcute Lymphoblastic Leukemia HL60 Cell Chronic Myelogenous Leukemia Acute Myelogenous Leukemia Terminal Deoxynucleotidyl Transferase
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