期刊
CYTOMETRY PART A
卷 69A, 期 12, 页码 1193-1201出版社
WILEY
DOI: 10.1002/cyto.a.20345
关键词
lactadherin; phosphatidylserine; apopitosis; leukemia; annexin V
Background: 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. Methods: We utilized fluorescein-labeled lactadherin together with the benchmark PS Probe, annexin V, to detect I'S 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 apopitosis. Results: 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 than or similar to 2.5%-8% for the membrane domains that stained with lactadherin but not annexin V. Conclusions: Macrophages may utilize lactadherin to detect PS exposure prior to exposure of sufficient PS to bind annexin V The methodology enables detection of PS exposure at earlier stages than established methodology. (c) 2006 International Society for Analytical Cytology
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据