Characterization of A Novel 65kDa E-selectin Ligand on G-CSF Mobilized Leukocytes

G-CSF 动员白细胞上新型 65kDa E-选择素配体的表征

基本信息

批准号：
7213644
负责人：
ROBERT SACKSTEIN
金额：
$ 26.25万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-07-01 至 2009-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7213644
关键词：
Acute Acute Lung Injury Adhesions Adhesives Affinity Antigens Arthritis Binding Biological Assay Biology Blood Vessels Bone Marrow Bone Marrow Cells Carbohydrates Cardiac Cell Adhesion Molecules Cell surface Cells Chimera organism Clinical Condition Dose E-Selectin Electrophoresis Epitopes Exhibits Flow Cytometry Gel Glycoproteins Granulocyte Colony-Stimulating Factor Harvest Hematopoietic Hematopoietic Stem Cell Mobilization Hematopoietic Stem Cell Transplantation Heterogeneity Human In Vitro Inflammatory Inflammatory Response Ischemia L-Selectin Lead Leukocytes Ligands Liquid substance Mass Spectrum Analysis Mediating Membrane Membrane Proteins Methods Modification Molecular Myeloid Cells Newborn Respiratory Distress Syndrome P-selectin ligand protein Pathologic Peripheral Blood Stem Cell Phase Physiological Preparation Proteins Range Relative (related person)Series Serum Source Time Vascular Endothelium Western Blotting base clinically relevant comparative glycosylation glycosyltransferase in vivo insight interest leukocyte mediator novel novel strategies peripheral blood prevent progenitor protein E research study

项目摘要

DESCRIPTION (provided by applicant): Granulocyte-colony stimulating factor (G-CSF)-mobilized peripheral blood (MPB) has replaced traditional bone marrow harvest as the preferred source of cells for hematopoietic stem cell transplantation. Though generally considered to be a safer alternative to harvest, there are increasing observations that G-CSF administration causes vascular and inflammatory complications due to enhanced adhesion of leukocytes to vascular endothelium. A major mediator of leukocyte-endothelial adhesive interactions is E-selectin, an inducible endothelial molecule that binds sialofucosylated carbohydrate antigens recognized by the mAb HECA-452. We reasoned that vascular and inflammatory complications may result from G-CSF-induced expression of E-selectin ligands on circulating leukocytes. We have found that P-selectin glycoprotein ligand-1 (PSGL-1) and Hematopoietic Cell E-and L-selectin Ligand (HCELL) are E-selectin ligands on G- CSF MPB leukocytes. Importantly, our studies also reveal that a novel HECA-452-reactive ~65kDa high affinity E-selectin ligand (hereinafter called "~65kDa protein") is expressed on G-CSF MPB leukocytes but not on native leukocytes and not on normal human bone marrow cells. This ~65kDa protein exhibits potent Ca2+-dependent E-selectin ligand activity, as evidenced by (i) binding to fluid phase E-selectin and (ii) avid binding of E-selectin- transfected cells under physiological shear flow conditions. Treatment of normal human bone marrow cells with G-CSF at a pharmacokinetically-relevant concentration in vitro induces the expression of this ~65kDa protein directly on myeloid cells. We hypothesize that this ~65kDa E-selectin ligand contributes to vascular and inflammatory complications following G-CSF administration. The specific aims of this proposal are: (1) To identify the ~65kDa protein serving as an E-selectin ligand on G-CSF MPB leukocytes; and (2) To identify the subset(s) of human leukocyte-series cells that express this glycoprotein following G-CSF administration in vivo and in vitro. The identification and the characterization of expression of this novel E-selectin ligand should provide insights into the biology of E-selectin ligands and greatly increase our understanding of G-CSF MPB leukocyte-endothelial interactions. It is anticipated that the results obtained will lead to new approaches for preventing or alleviating G-CSF-induced vascular and inflammatory complications

描述（由申请人提供）：粒细胞 - 刺激因子（G-CSF）锻造的外周血（MPB）已取代了传统的骨髓收获，成为造血干细胞移植的细胞首选。尽管通常认为是收获的更安全的替代品，但由于白细胞对血管内皮的粘附增强，越来越多的观察结果是，G-CSF给药会引起血管和炎症并发症。白细胞 - 内皮粘合剂相互作用的主要介体是E-选择素，这是一种可诱导的内皮分子，它结合了MAB HECA-452识别的唾液氟糖基化的碳水化合物抗原。我们认为，血管和炎症并发症可能是由于G-CSF诱导的E-选择蛋白配体在循环白细胞上的表达而引起的。我们发现，P-选择蛋白糖蛋白配体1（PSGL-1）和造血细胞E和L-选择蛋白配体（HCELL）是G-CSF MPB MPB MPB Leukocytes上的e-纤维蛋白配体。重要的是，我们的研究还表明，一种新型的HECA-452反应性〜65KDA高亲和力E-选择蛋白配体（以下称为“ 〜65KDA蛋白”）在G-CSF MPB MPB白细胞上表达，但在正常的人类骨骼上不受欢骨髓细胞。该〜65KDA蛋白表现出有效的Ca2+依赖性的E-选择蛋白配体活性，这是通过（i）与生理剪切流动条件下E-选择转染细胞的液相E-选择素结合和（ii）E-选择转染细胞的狂热结合所证明的。用G-CSF治疗正常的人骨髓细胞在药代代动力学与相关的体外浓度上，可直接在髓样细胞上诱导该〜65KDA蛋白的表达。我们假设这种〜65KDA E-选择蛋白配体在G-CSF给药后会导致血管和炎症并发症。该提案的具体目的是：（1）确定〜65KDA蛋白在G-CSF MPB白细胞上用作E-选择蛋白配体；（2）确定人类白细胞系列细胞的子集，这些细胞在体内和体外施用G-CSF后表达这种糖蛋白。这种新型的E-选择蛋白配体的表达的鉴定和表征应提供对E-选择蛋白配体生物学的见解，并大大增加了我们对G-CSF MPB白细胞 - 内皮相互作用的理解。预计获得的结果将导致预防或减轻G-CSF诱导的血管和炎症并发症的新方法