Exposure to phorbol diester (TPA) in vitro as an aid in the classification of blasts in human myelogenous and lymphoid leukemias: In vitro differentiation, growth patterns, and ultrastructural observations

Abstract

Leukemic cells from the peripheral blood of 52 patients with acute and chronic leukemias were incubated with 12-0-tetradecanoyl phorbol ester (TPA). Thirty-one cases of lymphocytic leukemia (18 cases of acute lymphoblastic and 13 cases of chronic lymphocytic leukemia), 13 cases of acute nonlymphoblastic (myelo or myelomonoblastic) leukemia, and eight cases of blastic crisis of CGL (seven cases of predominantly myeloblastic crisis, and one case of lymphoblastic crisis) were studied. In all cases of lymphoid leukemia, cells formed clumps or aggregates after exposure to TPA, while in all cases of myeloid leukemia cells became adherent to the substrate. Seven of the eight cases of blastic crisis of CGL were predominantly myeloid in type and cells adhered to the substrate, while in a single case of lymphoid crisis in CGL cells formed clumps after TPA exposure. Functional, cytochemical, and ultrastructural studies showed altered cell differentiation and continuing in vitro maturation of leukemic cells after exposure to TPA. In the light of the above results, it is concluded that this simple test employing TPA exposure in vitro serves as a reliable means of distinguishing blasts from different origins in human leukemias.

Induction of plasmacytoid and hairy cell features by phorbol esters (TPA) in B‐lymphoma cells: Attempted correlation with disease activity

“…SEM was performed as described in earlier studies (Gamliel et al, 1983;Polliack et al, 1982Polliack et al, , 1986a. Briefly, cells were initially fixed in suspension with 2 per cent glutaraldehyde in PBS for at least 1 h. Thereafter cells were prepared for SEM using the GTGO procedure as described in detail in an earlier study (Polliack et al, 1986a).…”

“…In the past TPA has been shown to induce differentiation in different human leukemic and non-malignant lymphoblastoid cell lines (Huberman and Callahan, 1979;Okamura et al, 1984;Roth et al, 1982;Sacchi et al, 1984). Furthermore, TPA has been produced in v i m differentiation of a variety of leukemic cells derived from patients with acute leukemia (Greenberger et al, 1980;Koeffler et al, 1980;Polliack et al, 1982), hairy cell leukemia (HCL) , and chronic lymphocytic leukemia (CLL) (Dufer and Bernard, 1984;Nilsson et al, 1985;Okamura et al, 1982;Polliack et al, 1986a;Ralph and Kishimoto, 1982;Totterman et al, 1980Totterman et al, , 1981. TPA-induced alterations in human CLL cells appears to be the most studied system, probably because of the easy access to large numbers of circulating cells in this disorder.…”

Myeloid leukemia differentiation by phorbol ester and retinoic acid: A practical approach

“…uration (10,13). RA, on the other hand, seems to induce maturation of acute promyelocytic leukemia (3), while other types of nonlymphocytic leukemia do not respond (8,9).…”

“…The development of a malignant phenotype may be viewed as a defect in the normal process of differentiation, and numerous efforts have been dedicated to the study of substances that can induce cell maturation, including phorbol diester (TPA) (l), retinoic acid (RA) (2), dimethylsulfoxide (DMSO) (3), and 1-a-25-dihydroxyvitamin D3 (4). Chemically induced differentiation has been studied primarily in cell lines (5,6), though study has been extended to a number of fresh tumors (7), including fresh leukemic cells (8)(9)(10)(11)(12)(13). Exposure of leukemic cells in vitro to various inducers was used to help reveal the process of differentiation (10,14,15> or to allow better classification of the type of cell involved ( 1 1, 13,16).…”

Cytochemistry and ultrastructure in lymphoma and leukemia: Utility in the diagnosis of different leukemias and the recognition of subtypes of lymphoproliferative disorders

“…This review is based on our experience with over 200 cases, studied during the past three years. The results of this experience have been published in detail during 1980-1982(Polliack. 1977: Polliack e f a l , 1983a: Gamliel etal., 1981: Gamliel and Polliack, 1981 and all techniques used are outlined in these publications and will not be detailed in this review.…”