Immunofluorescent and Biochemical Studies of Terminal Deoxynucleotidyl Transferase in Treated Acute Leukaemia

Abstract

Summary. Indirect Immunofluorescence (IF) for terminal deoxynucleotidyl trans-ferase (TdT) was used in conjunction with the biochemical assay of TdT enzymatic activity to study human leukaemias before and during therapy. In addition, non-leukaemic marrows were analysed to compare the enzyme expression on normal cells. An excellent correlation was observed between the IF and biochemical methods when specimens contained greater than 5% TdT⁺ cells (by IF); below this level the biochemical assay was less reliable, while the sensitive IF test could detect isolated TdT⁺ cells among greater than 10 000 TdT negative cells. The IF method also had the advantage of allowing further immunological characterization of TdT⁺ cells, by simultaneous labelling of membrane antigens with appropriate antiscra. TdT⁺ cells expressing la-like antigens (but lacking other antigens associated with B- and T- lymphoid differentiation) were frequently found in low numbers in remission marrows from acute lymphoblastic leukaemia (ALL) patients. However, similar cells were also observed in remission acute myeloid leukaemia, as well as in non-leukaemic regenerating marrows, and marrow from normal donors. The presence of these normal TdT⁺ precursor cells therefore precluded the use of either IF or biochemical TdT tests for estimating the degree of residual disease or predicting early relapse in patients with non-T, non-B ALL. In contrast, the detection of TdT⁺ cells with T lymphoid antigens (HuTLA⁺) but lacking Ia antigens, in thymic (T-cell) -ALL, but not in normal marrow, allowed the use of this combination of markers to detect minimal residual disease in T-ALL.