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Specific detection of monocytic lysozyme within normal and leukemic cells
C Leculier1, N Couprie, A Francina
1Unité d'Immunologic, Institut Pasteur de Lyon, France.
Blood
|February 1, 1992
Summary
A new monoclonal antibody specifically targets human lysozyme in monocytes, aiding in acute myeloid leukemia subtype differentiation. This antibody also inhibits lysozyme
Area of Science:
- Immunology
- Hematology
- Biochemistry
Background:
- Lysozyme is an enzyme found in various immune cells.
- Distinguishing between normal and leukemic monocytes is crucial for diagnosis.
- Monocyte-specific markers are valuable for hematological research.
Purpose of the Study:
- To develop and characterize a murine monoclonal antibody against human lysozyme (AHL MoAb).
- To investigate the expression of lysozyme in normal and leukemic monocytes.
- To assess the potential of AHL MoAb in differentiating acute myeloid leukemia subtypes.
Main Methods:
- Production of a murine monoclonal antibody against human lysozyme.
- Flow cytometry analysis of normal monocytes and neutrophils.
- Immunoblotting of monocyte and polymorphonuclear leukocyte (PMNL) lysates.
- Enzymatic activity assay using lysozyme substrate.
- Flow cytometric analysis of leukemic cells from acute myeloid leukemia (AML) patients (FAB M1-5).
Main Results:
- AHL MoAb demonstrated specific reactivity with normal monocytes (82%-98% positive) but not neutrophils.
- Immunoblotting confirmed lysozyme presence in both monocyte and PMNL lysates.
- AHL MoAb significantly inhibited lysozyme enzymatic activity.
- Leukemic monocytes from FAB M5 subtype showed significantly higher lysozyme positivity compared to other AML subtypes.
Conclusions:
- The AHL MoAb recognizes a lysozyme epitope near the catalytic site, predominantly expressed on monocytes.
- This epitope appears masked in polymorphonuclear leukocyte (PMNL) granules.
- AHL MoAb shows promise as a diagnostic tool for differentiating acute myeloid leukemia subtypes, particularly FAB M5.