Monocyte-selective transendothelial migration: dissection of the binding and transmigration phases by an in vitro assay.
Monocyte-selective transendothelial migration: dissection of the binding and transmigration phases by an in vitro assay.
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DOI：
10.1084/jem.176.3.819
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发表时间：
1992-09-01
期刊：
影响因子：
--
通讯作者：
Weigl SA
中科院分区：
文献类型：
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作者：
Muller WA;Weigl SA
我们描述了跨内皮迁移（TEM）的定量测定法，该测定使我们能够选择性地研究单核细胞与汇合的人内皮细胞（HEC）单层的相互作用。 HEC在水合胶原蛋白凝胶上生长；单核细胞无需纯化。 100％的单核细胞在37摄氏度的1小时内将单层转移，并积聚在内皮下胶原蛋白中；在这段时间内未检测到淋巴细胞的TEM。中性粒细胞从同一供体的迁移速度较慢且不完整，只有14％的PMN在2小时内迁移。这种快速的TEM发生在没有外源性趋化剂的情况下，并且该系统中的HEC不会表达细胞因子诱导的白细胞粘附分子。对TEM测定法的稍作修改使我们能够将与顶端HEC表面的结合与TEM分离。我们发现紧密的顶表结合是TEM的速率限制步骤。这种结合和TEM的三分之二可以通过针对白细胞β2整合蛋白链CD18的单克隆抗体阻止。该测定法可以使我们能够剖析二尿的结合和移民阶段的机制。
We describe a quantitative assay of transendothelial migration (TEM) that allows us to selectively study the interaction of monocytes with confluent human endothelial cell (HEC) monolayers. The HEC are grown on hydrated collagen gels; the monocytes need not be purified. 100% of monocytes transmigrated the monolayer within 1 h at 37 degrees C and accumulated in the subendothelial collagen; TEM of lymphocytes was not detected within this time. Migration of neutrophils from the same donor was much slower and incomplete, with only 14% of PMN transmigrating in 2 h. This rapid TEM occurs in the absence of exogenous chemoattractants, and HEC in this system do not express cytokine- inducible leukocyte adhesion molecules. A slight modification of the TEM assay allowed us to separate binding to the apical HEC surface from TEM. We found that tight apical surface binding was the rate-limiting step for TEM. Two-thirds of this binding and TEM could be blocked by a monoclonal antibody against the leukocyte beta 2 integrin chain CD18. This assay will allow us to dissect the mechanisms of both the binding and transmigration stages of diapedesis.