title: Expression of P-glycoprotein and multidrug resistance-associated protein in healthy volunteers and HIV-infected patients.
abstract: Increased expression of the multidrug efflux transporters P-glycoprotein (P-gp) and multidrug resistance-associated protein (MRP) has been suggested as a potential mechanism for decreased drug availability at certain intracellular sites that provide sanctuary for HIV. Here we investigate the expression of these transporters in peripheral blood mononuclear cells (PBMCs) of HIV-infected patients and healthy volunteers. Venous blood (30 ml) was taken from healthy volunteers (n = 21) and HIV-infected patients (n = 21; 4 antiretroviral drug naive, 17 antiretroviral drug experienced). PBMCs were isolated and fixed. To assess P-gp expression, PBMCs were incubated with an isotype control antibody or an antibody directed to an external epitope of P-gp (UIC2). To assess MRP expression, cells were permeabilized before incubation with either a control antibody or an antibody directed to an internal epitope of MRP (MRPm5). After washing, a secondary phycoerythrin-bound antibody was incubated. After additional wash steps, samples were fixed and analyzed by flow cytometry. The median fluorescence intensity of 5000 events was recorded. Results are expressed as fold increase between isotype control and UIC2/MRPm5 samples. Expression of P-gp in HIV-infected patients (1.42 +/- 0.36) was significantly lower (p = 0.0021; 95% CI, -0.633 to -0.164) than in healthy volunteers (1.82 +/- 0.55). However, MRP expression was similar in HIV-infected patients (1.37 +/- 0.34) and healthy volunteers (1.37 +/- 0.21; p = 0.91; 95% CI, -0.148941 to 0.165191). We conclude that in HIV infection, P-gp expression in total PBMCs is reduced whereas MRP expression appears to be unaltered.
authors.affiliation: Department of Pharmacology and Therapeutics, University of Liverpool, New Medical Building, Ashton Street, Liverpool L69 3GE, UK. ermeaden@liverpool.ac.uk
authors.forename: E R
authors.initials: ER
authors.lastname: Meaden
authors.forename: P G
authors.initials: PG
authors.lastname: Hoggard
authors.forename: B
authors.initials: B
authors.lastname: Maher
authors.forename: S H
authors.initials: SH
authors.lastname: Khoo
authors.forename: D J
authors.initials: DJ
authors.lastname: Back
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000096:biosynthesis / Q000097:blood*; D000328:Adult; D005260:Female; D015658:HIV Infections / Q000097:blood*; D015497:HIV-1; D006801:Humans; D016130:Immunophenotyping; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D008297:Male; D008856:Microscopy, Fluorescence; D008875:Middle Aged
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1