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Isolation of Human Monocytes by Double Gradient Centrifugation and Their Differentiation to Macrophages in Teflon-coated Cell Culture Bags
Published on: September 9, 2014
Detection of a monocyte/macrophage differentiation antigen in routinely processed paraffin-embedded tissues by
H J Radzun1, M L Hansmann, H J Heidebrecht
1Institute of Pathology, University of Kiel, Germany.
Insights
A novel monoclonal antibody, Ki-M1P, specifically targets monocytes and macrophages. This antibody is a valuable diagnostic tool for identifying monocytic leukemias and studying the human monocyte/macrophage system.
Area of Science:
- Immunology
- Hematology
- Cell Biology
Background:
- Monocytes and macrophages are crucial immune cells involved in various physiological and pathological processes.
- Accurate identification and characterization of these cells are essential for diagnosing hematological malignancies and understanding immune responses.
Purpose of the Study:
- To describe a new monoclonal antibody, Ki-M1P, for recognizing human monocytes and their macrophage derivatives.
- To evaluate the utility of Ki-M1P as a diagnostic marker in hematopathology.
Main Methods:
- Development of monoclonal antibody Ki-M1P against human lymph node tissue.
- Immunohistochemical analysis (light and electron microscopy) to determine antigen specificity.
- Western blot analysis to characterize the molecular weight of the recognized antigen.
Main Results:
- Ki-M1P specifically recognizes monocytes and macrophages, with no reactivity observed in granulocytes, dendritic cells, or other cell types.
- The antibody is effective in distinguishing monocytic leukemias (FAB M4 and M5).
- The recognized antigen comprises multiple proteins in blood monocytes but primarily a 60 kDa protein in tissue macrophages.
Conclusions:
- Ki-M1P is a specific and reliable marker for the human monocyte/macrophage system.
- Its stability during routine tissue processing makes it a valuable reagent for retrospective diagnostic studies.
- Ki-M1P aids in the classification of monocytic leukemias and the study of monocyte/macrophage system disorders.
Abstract:
A new monoclonal antibody Ki-M1P that is raised against supernatants of detergent solubilized human lymph node tissue is described. Ki-M1P recognizes in particular monocytes and their macrophage derivatives as tested by light- and electron-microscopic immunohistochemistry. Granulocytes, dendritic cells as the accessory cells of humoral and cellular immune response, and epithelial, endothelial, neural, and mesenchymal cells do not react with Ki-M1P. In extensive application Ki-M1P has proven to be a useful marker for distinguishing monocytic leukemias within FAB groups M4 and M5. The recognized antigen is composed of five proteins with molecular masses of about 60, 92, 98, 124, and 150 kDa in blood monocytes, whereas tissue macrophages tested so far expressed only the 60-kDa protein. Because the Ki-M1P antigen is not destroyed or masked during routine fixation and paraffin embedding of biopsy tissue samples, Ki-M1P represents a useful diagnostic reagent for the identification of physiological functional and pathologic reaction forms as well as neoplastic variants of the human monocyte/macrophage system even in retrospective studies.
