使用UIC2迁移分析对人外周血淋巴细胞中P-糖蛋白介导的膜转运进行分析。

Analysis of P-glycoprotein-mediated membrane transport in human peripheral blood lymphocytes using the UIC2 shift assay.

作者信息

Park Suk W, Lomri Noureddine, Simeoni Luiz A, Fruehauf John P, Mechetner Eugene

机构信息

Ingenex, Inc., Menlo Park, California, USA.

出版信息

Cytometry A. 2003 Jun;53(2):67-78. doi: 10.1002/cyto.a.10039.

DOI:10.1002/cyto.a.10039

PMID:12766968

Abstract

BACKGROUND

During transport-associated adenosine triphosphate hydrolysis, P-glycoprotein (Pgp) undergoes conformation transitions detected by UIC2, a functional anti-Pgp monoclonal antibody. A newly developed UIC2 shift assay is based on increased UIC2 reactivity in the presence of Pgp substrates. All peripheral blood leukocytes express low Pgp levels. The existing antibody-based detection methods are limited in their sensitivity and require additional techniques to simultaneously analyze Pgp expression and efflux, making it difficult to ascertain the physiologic role of Pgp-mediated transport.

METHODS

We validated the UIC2 shift assay against UIC2 immunostaining and DiOC(2) efflux. The UIC2 shift assay was then used to characterize Pgp functional expression and its physiologic substrates in peripheral blood leukocytes.

RESULTS

A strong correlation was observed between the UIC2 shift assay versus immunostaining and dye efflux tests. The UIC2 shift assay showed improved sensitivity (compared with conventional UIC2 staining) and allowed for simultaneous detection of Pgp expression and function. Using this assay, we identified several new Pgp substrates, including monensin and retinol, and confirmed that interleukin-2 and interferon-gamma can be transported by Pgp.

CONCLUSIONS

Our findings validate the use of the UIC2 shift assay in MDR1 detection and support the idea that Pgp plays a physiologic role in immunoregulation.

摘要

背景

在与转运相关的三磷酸腺苷水解过程中，P-糖蛋白（Pgp）会发生构象转变，可被功能性抗Pgp单克隆抗体UIC2检测到。一种新开发的UIC2位移测定法基于在Pgp底物存在时UIC2反应性的增加。所有外周血白细胞表达的Pgp水平较低。现有的基于抗体的检测方法灵敏度有限，需要额外技术来同时分析Pgp表达和外排，这使得难以确定Pgp介导的转运的生理作用。