Monoclonal antibodies against human granulocytes and myeloid differentiation antigens |
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Authors: | Patrice Mannoni Anna Janowska-Wieczorek A Robert Turner Locksley McGann Jean-Michael Turc |
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Institution: | a Canadian Red Cross, Blood Transfusion Service and University of Alberta. Department of Pathology, Edmonton, Canada b Department of Medicine, Cross Center Institute, Edmonton, Alberta, Canada |
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Abstract: | Monoclonal antibodies (MCA) were obtained by immunizing BALB/c mice with 99% pure granulocytes from normal donors or with a whole leukocyte suspension obtained from a chronic myelogenous leukemia (CML) patient, and then fusing the mouse spleen cells with a 315–43 myeloma cell clone. Four MCA were selected and studied using ELISA, immunofluorescence, cytotoxicity assays, and FACS analysis. Antibodies 80H.1. 80H.3. and 80H.5 (from normals) and 81H.1 (from CML) detected antigens expressed on neutrophils. Antibodies 80H.1 and 80H.3 (lgG) also reacted with monocytes but not with other blood cell subsets. Antibodies 80H.5 and 81H.1 (lgM) were cytotoxic and reacted strongly with most of the cells of the neutrophil maturation sequence. i.e., myeloblasts, promyelocytes, myelocytes, and mature granulocytes. Antibodies 80H.5 and 81H.1 also inhibited BFU-GM and CFU-E. Antigens recognized by 80H.3. 80H.5, and 81H.1 were expressed both on a proportion of cells from HL.60, KG.1, ML.1, and K562 myeloid cell lines, and on a proportion of blast cells isolated from patients with acute myelogenous leukemia. They were not found on lymphoid cell lines or lymphoid leukemia cells. These MCA recognize either late differentiation antigens expressed on mature neutrophils and monocytes (80H.1 and 80H.3) or early differentiation antigens (80H.5 and 81H.1) specific to the granulocytic lineage. They may be useful for a better definition of those antigens specific to hematopoietic stem cells and their relationship with normal or neoplastic hematopoiesis. |
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Keywords: | ALL acute lymphocytic leukemia ELISA enzyme-linked immunosorbent assay AML acute myelocytic leukemia BFU-E burst forming unit Erythoroid FACS fluorescent activated cell soreter BSA bovine serum albumin FITC fluorescein isothiocyanate CFU-GM colony forming unit granulocyte/macrophage GAT granuloaalutination test GCT granulocytotoxicity test CFU-E colony forming unit erythroid HAT hypoxanthine-aminopterine-thymidine CML chronic myelocytic leukemia IFT immunofluorescence test MCA monoclonal antibodies FCS fetal calf serum PBS phosphate buffer saline |
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