Nov 24, 2006

Lactadherin detects early phosphatidylserine exposure on immortalized leukemia cells undergoing programmed cell death

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
J ShiGary E Gilbert


Phosphatidylserine (PS) appears on the outer membrane leaflet of cells undergoing programmed cell death and marks those cells for clearance by macrophages. Macrophages secrete lactadherin, a PS-binding protein, which tethers apoptotic cells to macrophage integrins. We utilized fluorescein-labeled lactadherin together with the benchmark PS Probe, annexin V, to detect PS exposure by flow cytometry and confocal microscopy. Immortalized leukemia cells were treated with etoposide, and the kinetics and topology of PS exposure were followed over the course of apoptosis. Costaining etoposide-treated leukemoid cells with lactadherin and annexin V indicated progressive PS exposure with dim, intermediate, and bright staining. Confocal microscopy revealed localized plasma membrane staining, then diffuse dim staining by lactadherin prior to bright generalized staining with both proteins. Annexin V was primarily localized to internal cell bodies at early stages but stained the plasma membrane at the late stage. Calibration studies suggested a PS content less, less than or approximately equal to 2.5%-8% for the membrane domains that stained with lactadherin but not annexin V. Macrophages may utilize lactadherin to detect PS exposure prior to ...Continue Reading

Mentioned in this Paper

Flow Cytometry
Annexin A5
Apoptosis, Intrinsic Pathway
Outer Membrane
Leukemoid Reaction

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