The Use of Monoclonal Antibodies Against Primary Myeloid Granules in Normal and Leukemic Cells

Abstract

Three monoclonal antibodies, K101, D46, and H36/71 (CD15), reactive with membrane components of primary granules of human promyelocytes, were studied to assess their binding to normal and leukemic cells. Using the alkaline phosphatase antialkaline phosphatase technique, these antibodies were applied to sections of normal organs and to peripheral blood and bone marrow films from hematologically normal individuals and patients with hematologic malignancies. In control experiments, antibodies showed reactivity with cytoplasmic constituents of granulocytes from the promyelocytic to the neutrophilic stage. In acute myeloid leukemia, antibody K101 was positive (more than 20% of blasts) in 13 of 21 (62%) cases, while antibody D46 was positive in 11 of 17 (65%) cases. Antibody H36/71 was positive in only 4 of 24 (17%) cases of acute myeloid leukemia. At least one marker was present in 6 of 8 (75%) cases of acute lymphoblastic leukemia with myeloid antigen-positive blasts and was negative in 20 cases of acute lymphoblastic leukemia with myeloid antigen-negative blasts. These results support the view that abnormal granules (with defective expression of the D46, K101, and H36/71 antigens) form in blastic and leukemic cells of patients with acute myeloid leukemia. Data also suggest that membrane components of myeloid granules are made in the cytoplasm of cells from some acute lymphoblastic leukemia patients with myeloid antigen-positive blasts.

Double Esterase Staining andOther Neutrophilic Granule Abnormalities in 237 Patients with the Myelodysplastic Syndrome Studied by the Cancer and Leukemia Group B

“…Another monoclonal antibody H36/71, prepared against the HL60 cell line, was provided by one of us (A.K.S.). This antibody recognizes a CD-15-related glycoprotein epitope (3-fucosyl-N-acetyllactosamine) present on the neutrophilic cell surface and on both primary and secondary neutrophilic granule membranes [8]. However, following tissue fixation, this antibody reacts mostly with neutrophilic granule membranes.…”

“…However, following tissue fixation, this antibody reacts mostly with neutrophilic granule membranes. All immunocytochemical studies were carried out using an alkaline phosphatase antialkaline phosphatase method as previously described [4, 8]. …”

Surface Antigen Changes during Normal Neutrophilic Development: A Critical Review

“…CD15s expression was downregulated then was upregulated with terminal differentiation (28). CD15 has an intracellular pool in the membranes of the primary and secondary granules, and surface expression is increased upon stimulation with chemotactic peptides (46,55). CD15s is increased on PB neutrophils with G-CSF administration (56).…”