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

摘要: 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 reactivity in the presence of Pgp substrates. All peripheral blood leukocytes express low levels. The existing antibody-based detection methods are limited their sensitivity and require additional techniques to simultaneously analyze expression efflux, making it difficult ascertain physiologic role Pgp-mediated transport. Methods We validated against immunostaining DiOC2 efflux. was then used characterize its substrates leukocytes. Results A strong correlation observed between versus dye efflux tests. showed improved (compared with conventional staining) allowed for simultaneous function. Using this assay, we identified several new substrates, including monensin retinol, confirmed that interleukin-2 interferon-γ can be transported Pgp. Conclusions Our findings validate use MDR1 support idea plays immunoregulation. Cytometry Part 53A:67–78, 2003. © 2003 Wiley-Liss, Inc.