Comparison of two methods to detect P-glycoprotein in patients with chronic lymphocytic leukaemia.
P-glycoprotein (Pgp) is a transmembrane protein associated with multiple drug resistance. Pgp can be detected by several monoclonal antibodies or its activity inferred by measuring drug uptake. We compared two methods for quantitating Pgp in 32 patients with chronic lymphocytic leukaemia. The monoclonal antibody 4E3, which recognizes an external epitope of Pgp, was detected by flow cytometry. Intracellular daunorubicin (DNR) accumulation was measured by flow cytometry in the presence (treated) and absence (control) of cyclosporin, an agent known to inhibit Pgp. Correlation between the degree of positivity on the drug uptake assay and Pgp detected by monoclonal antibody 4E3 was poor (r = 0.06). No association with previous drug exposure or lymphocyte doubling time and Pgp positivity was found in this series of patients. Poor correlation between assays might reflect a lack of sensitivity of the DNR uptake assay. Drug accumulation may be influenced by other cellular efflux pumps unrelated to Pgp, making the DNR assay non-specific.
Chin-Yee
I
I
Flow Cytometry Unit, London Health Sciences Centre, University of Western Ontario, Canada.
Alshammari
S
S
Anderson
L
L
Kadri
M
M
Keeney
M
M
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000276:immunology; D000328:Adult; D000368:Aged; D000911:Antibodies, Monoclonal / Q000276:immunology; D002462:Cell Membrane / Q000737:chemistry; D016572:Cyclosporine / Q000494:pharmacology; D003630:Daunorubicin / Q000378:metabolism; D018432:Drug Resistance, Multiple; D019008:Drug Resistance, Neoplasm; D000939:Epitopes / Q000276:immunology; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000378:metabolism* / Q000473:pathology; D008875:Middle Aged; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000911:Antibodies, Monoclonal; D000939:Epitopes; D016572:Cyclosporine; D003630:Daunorubicin