ADHESION MOLECULE EXPRESSION DURING NEUTROPHIL CHEMOTAXIS

中性粒细胞趋化过程中粘附分子的表达

• 批准号: 5200847
• 负责人: L HARVATH
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/5200847
• 关键词: CD antigens; antibody receptor; cell adhesion molecules; cell migration; cell population study; chemoattractants; chemotaxis; flow cytometry; human tissue; inflammation; neutrophil; receptor expression

The goal of this project is to determine whether human neutrophil (PMN) surface antigen expression is altered after PMN have migrated towards an inflammatory stimulus. We have previously shown that PMN adhesion molecule expression is differentially downregulated after PMN chemotaxis. The present study was undertaken to examine the changes that occur in PMN surface molecule expression of aminopeptidase N (CD13) and the Fcg receptors, FcRII (CD32) and FcRIII (CD16), after PMN chemotaxis in vitro. Two populations of PMN are easily identified and separated with an in vitro polycarbonate membrane chemotaxis system; one population does not migrate towards chemoattractant (nonmigrating population) and remains on the upper surface of the membrane, whereas the other population migrates through the membrane pores to the lower surface of the membrane (migrating population). PMN, incubated in suspension with or without the N-formyl peptide chemoattractant (FMLP), were compared with the migrating and nonmigrating subpopulations which were exposed to a gradient of FMLP. PMN were stained with a panel of monoclonal antibodies which recognize aminopeptidase N (CD13), or the Fcg receptors, FcRII (CD32) and FcRIII (CD16), and surface expression was quantified with a flow cytometer. CD13 expression was not significantly altered after PMN chemotaxis. CD13 expression on migrating and non-migrating PMN populations was equivalent. In contrast, FcRII and FcRIII expression was down- regulated on the migrating PMN subpopulation by 42% and 58%, respectively, when compared with the non- migrating PMN subpopulation. Down-regulation of CD32 did not occur when non-adherent PMN were stimulated with chemoattractant in suspension, however, CD16 expression was down-regulated by 42% when non-adherent PMN were stimulated with FMLP in suspension. The results of this study demonstrate that PMN surface molecule expression is selectively down-regulated after PMN chemotaxis. This experimental system provides a means for quantifying the expression of myeloid antigens which are important in inflammatory responses.

本项目的目标是确定人类中性粒细胞（PMN） 当中性粒细胞向中性粒细胞表面迁移后， 炎症刺激。 我们之前已经证明， PMN趋化后分子表达差异下调。 本研究的目的是探讨中性粒细胞的变化， 氨基肽酶N（CD 13）和Fcg的表面分子表达 受体，FcRII（CD 32）和FcRIII（CD 16），在体外PMN趋化后。 两个群体的中性粒细胞很容易识别和分离， 体外聚碳酸酯膜趋化系统;一个群体不 向化学引诱物迁移（非迁移种群）并保持在 膜的上表面，而其他种群则迁移 通过膜孔到达膜的下表面 （流动人口）。 PMN，在有或没有 N-甲酰肽化学引诱物（FMLP）与迁移性 和非迁移亚群，暴露于梯度的FMLP。 PMN用一组单克隆抗体染色， 氨肽酶N（CD 13）或Fcg受体FcRII（CD 32）和FcRIII （CD 16），并用流式细胞仪定量表面表达。 PMN趋化后CD 13表达无明显改变。 CD13 迁移性和非迁移性中性粒细胞群的表达是相等的。 相反，FcRII和FcRIII的表达下调， 迁移性PMN亚群分别减少42%和58%， 非迁移性中性粒细胞亚群。 CD 32的下调 当非粘附的PMN被化学引诱物刺激时， 然而，在悬浮液中，CD 16表达下调42%， 用悬浮的FMLP刺激非粘附的PMN。 本研究结果表明，PMN表面分子 表达在PMN趋化后选择性下调。 这 实验系统提供了一种用于定量表达 在炎症反应中重要的髓样抗原。