Characteristics of multidrug resistance in Plasmodium and Leishmania: detection of P-glycoprotein-like components.
Multidrug-resistance (MDR) in neoplastic cells is frequently characterized by the overexpression of P-glycoprotein (PGP), a 170 kDa transmembrane glycoprotein that binds multiple cytotoxic drugs as well as calcium channel antagonists. Chloroquine resistance in Plasmodium falciparum appears to be analogous to MDR in neoplastic cells, where the induction of resistance with one drug confers resistance to other structurally and functionally unrelated drugs. To test the hypothesis that chloroquine resistance in P. falciparum and antimony resistance in Leishmania is mediated by a similar mechanism of MDR in mammalian neoplastic cells, a PGP-specific monoclonal antibody (C219) was used to determine the presence of PGP genes in resistant and sensitive Plasmodium and Leishmania parasites by indirect immunofluorescence assays and Western blotting procedures. These PGP-like components were detected in both drug-sensitive and -resistant Plasmodium and Leishmania cells. A 40-42 kDa component was observed to be greater in a chloroquine-resistant P. berghei (C line) than in a chloroquine-susceptible P line. Differences observed between Pentostam-resistant and -sensitive Leishmania promastigote clones and isolates included the increased expression of 96-106 and 23-25 kDa peptides in drug-resistant L. enrietti, and increased amounts of two different peptides in two drug-resistant L. panamensis clones (i.e., 96-106 and 43-45 kDa in WR-746-CL4, and 53 and 23-25 kDa in kDa) in amastigotes as in MDR KB carcinoma cells (KB-V1). Comparative indirect immunofluorescent studies suggested that a correlation existed between the degree of antimony susceptibility and the concentration of the moiety recognized by C219 in two L. panamensis clones. Binding of the C219 monoclonal antibody to the PGP-like component of Leishmania was blocked by Pentostam, while the binding of C219 to multiple-drug resistant KB-V1 PGP was not inhibited by Pentostam, regardless of the PGP concentration. This suggests some degree of specificity in the binding of Pentostam to the Leishmania PGP-like components. In addition, these studies have demonstrated that drug-sensitive Leishmania accumulate two to five times more 125Sb-Pentostam than resistant clones.
Division of Experimental Therapeutics, Walter Reed Army Institute of Research, Washington, DC.
M
M
Grogl
R K
RK
Martin
A M
AM
Oduola
W K
WK
Milhous
D E
DE
Kyle
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000818:Animals; D000962:Antimalarials / Q000494:pharmacology*; D000967:Antimony Sodium Gluconate / Q000494:pharmacology; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D015153:Blotting, Western; D004351:Drug Resistance; D007891:Leishmania / Q000032:analysis / Q000187:drug effects*; D007892:Leishmania braziliensis / Q000032:analysis / Q000187:drug effects; D007894:Leishmania mexicana / Q000032:analysis / Q000187:drug effects; D008562:Membrane Glycoproteins / Q000737:chemistry*; D010961:Plasmodium / Q000032:analysis / Q000187:drug effects*; D010962:Plasmodium berghei / Q000032:analysis / Q000187:drug effects; D010963:Plasmodium falciparum / Q000032:analysis / Q000187:drug effects; D014407:Tumor Cells, Cultured / Q000737:chemistry
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000962:Antimalarials; D000981:Antiprotozoal Agents; D008562:Membrane Glycoproteins; D000967:Antimony Sodium Gluconate