The identification and characterization of umbilical cord blood-derived human basophils
The identification and characterization of umbilical cord blood-derived human basophils
复制标题
DOI：
10.1002/jlb.64.4.474
复制
发表时间：
1998-10-01
影响因子：
5.5
通讯作者：
Oliver, JM
中科院分区：
文献类型：
--
作者：
Kepley, CL;Pfeiffer, JR;Oliver, JM
人外周血嗜碱性粒细胞上的过敏原特异性免疫球蛋白 E 交联会导致组胺和其他炎症介质的释放，从而引发过敏和哮喘。从 IgE 结合到介质释放的信号通路尚未完全建立，主要是由于难以获得足够数量的高度纯化的嗜碱性粒细胞，本研究的目标是轻松获得高产量和高纯度的 Fc ε RI 阳性人嗜碱性粒细胞，用于信号转导途径的研究。我们描述了所有体外培养系统，其中用白细胞介素 3 (IL-3) 脉冲正常人脐带血白细胞 3-4 小时，然后在含有胎牛血清的培养基中孵育，培养 14 至 28 天期间，产生的细胞群主要为 Fc epsilon RI 阳性。当通过光学和电子显微镜检查时，这些细胞类似于外周血嗜碱性粒细胞。与正常血液嗜碱性粒细胞一样，它们表达整合素 CD11b、CD18、CD29 和 CD49d。大多数 IL-3 脉冲细胞还表达由嗜碱性粒细胞特异性抗体 2D7 识别的标记物，Fc epsilon RI 交联导致组胺的时间和剂量依赖性释放。 Fc epsilon RI 交联还会刺激蛋白质酪氨酸磷酸化，这被认为是导致 IgE 介导的外周血嗜碱性粒细胞激活的第一个事件。这些研究将脐带血确定为一种可利用的来源，可从中培养嗜碱性粒细胞样细胞来检查 Fc epsilon RI 介导的信号转导。
Cross-linking allergen-specific immunoglobin E on human peripheral blood basophils results in the release of histamine and other inflammatory mediators that initiate allergy and asthma. The signaling pathways leading from IgE binding to mediator release have not been well established, mainly due to the difficulty in obtaining adequate numbers of highly purified basophils, It was the goal of this study to easily obtain Fc epsilon RI-positive human basoplrils in high yield and purity for studies of signal transduction pathways, We describe all in vitro culture system in which pulsing normal human cord blood leukocytes with interleukin-3 (IL-3) for 3-4 h followed by incubation in medium with fetal bovine serum generates a cell population that is predominately Fc epsilon RI positive between 14 and 28 days of culture. These cells resemble peripheral blood basophils when examined by light and electron microscopy. Like normal blood basophils, they express the integrins, CD11b, CD18, CD29, and CD49d. A majority of the IL-3-pulsed cells also express a marker recognized by the basophil-specific antibody, 2D7, Fc epsilon RI cross-linking results in a time and dose-dependent release of histamine. Fc epsilon RI cross-linking also stimulates protein-tyrosine phosphorylation, thought to be the first event leading to the IgE-mediated activation of peripheral blood basophils, These studies establish cord blood as an accessible source from which basophil-like cells call be developed to examine Fc epsilon RI-mediated signal transduction.