Cellular efflux and uptake transports of several anti-HIV agents are mediated by plasma membrane-localized solute transporters. However, transporters involved in raltegravir disposition have not been fully characterized. Here, we performed in vitro studies to identify transporters mediating transcellular transport of raltegravir. Transepithelial raltegravir transport was examined using porcine kidney epithelial cell line (LLC-PK1) and LLC-PK1 cells stably transfected with P-glycoprotein (also known as Multiple drug resistance I) (L-MDR1). Transepithelial transport of raltegravir in Caco-2 cell monolayers, and intracellular accumulation of raltegravir in the MT-2 and MT-4 (CD4+ T-) cells were measured in the presence or absence of anti-HIV agents. The uptake of raltegravir was investigated in HEK-293 cells expressing each of several solute carrier family transporters, The apical-to-basal raltegravir transport was significantly decreased in L-MDR1 as compared to that in LLC-PK1 monolayers. In HEK-293 cells expressing breast cancer resistance protein (BCRP), raltegravir accumulation was lower than that in the mock-transfected cells. In Caco-2 cells, protease inhibitors including nelfinavir, ritonavir and lopinavir enhanced the apical-to-basal transport of raltegravir. By contrast, reverse transcriptase inhibitors such as zidovudine, efavirenz, and nevirapine, had no effect on raltegravir transport. The cellular accumulation of raltegravir in MT-2 cells, which express P-glycoprotein, was significantly increased in the presence of protease inhibitors. By contrast, protease inhibitors only marginally increased the accumulation of raltegravir in MT-4 cells, in which P-glycoprotein is not expressed. The present findings suggest that raltegravir is a substrate of both P-glycoprotein and BCRP. Protease inhibitors increase the absorptive transport of raltegravir in Caco-2 cells, and the cellular accumulation in T-cells, at least in part, by P-glycoprotein-mediated interaction. (C) 2013 Elsevier Inc. All rights reserved.