Characterization of Haemonchus contortus P-glycoprotein-16 and its interaction with the macrocyclic lactone anthelmintics

• Published in: Molecular and biochemical parasitology Vol. 204; no. 1; pp. 11 - 15
• Main Authors: Godoy, P., Che, H., Beech, R.N., Prichard, R.K.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01-11-2015
• Subjects: Abamectin; Animals; Antinematodal Agents - pharmacology; ATP Binding Cassette Transporter, Sub-Family B - genetics; ATP Binding Cassette Transporter, Sub-Family B - metabolism; Drug Resistance; Fluorescent Dyes - metabolism; Haemonchus - drug effects; Haemonchus - metabolism; Helminth Proteins - genetics; Helminth Proteins - metabolism; Ivermectin; LLC-PK1 Cells; Macrocyclic lactone resistance; Macrolides - pharmacology; Moxidectin; Nematode; P-glycoprotein; Protein Transport - drug effects; Rhodamine 123 - metabolism; Swine; mM; EPRI; GluCl; RNA; DMSO; RNB; Macrocyclic lactone resistance; μM; NBD; G418; RT-PCR; Nematode; cDNA; MDR; logp; ML; PBS; PCR;SDS; Abamectin; MO; DOR; qRT-PCR; MW; VSP; PMSF; RKP; CAT; FCS; PGP; λ; SEL; GAPDH; PCR; Hco-PGP-2; Ivermectin; ABA; IVM; C; kDa; mRNA; MRP; ABCtransporters; C-13, C-23, C-25; BZ; SOPs; Rho123; mdr1a; h; TMD; Moxidectin; m; Hco-Pgp-16; P-glycoprotein; DNA; MOX; PG; HBSS; HC; LEV; ATP
• ISSN: 0166-6851; 1872-9428
• DOI: 10.1016/j.molbiopara.2015.12.001

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•Hco-PGP-16 was expressed in mammalian LLC-PK1 cells.•Hco-PGP-16 translocated the fluorophore Rhodamine 123 (Rho123).•Avermectins, ivermectin and abamectin blocked Rho123 efflux by Hco-PGP-16.•Moxidectin produced an unsaturable, lesser effect on Rho123 transport by Hco-PGP-16.
Anthelmintic resistance in veterinary nematodes, including Haemonchus contortus, has become a limitation to maintaining high standards of animal health. Resistance in this parasite, to all drug families including the macrocyclic lactones (MLs) is a serious issue worldwide. Mechanisms of resistance to the MLs appear to be complex and to include the elimination of these compounds by ABC transporter-like proteins present in nematodes. In order to investigate the potential involvement of ABC transporters in ML resistance in H. contortus, we have characterized the functionality of the ABC transporter H. contortus P-glycoprotein-16 (Hco-PGP-16) expressed in mammalian cells. This has included a study of its interaction with different MLs, including the avermectins, abamectin (ABA) and ivermectin (IVM), and the milbemycin, moxidectin (MOX). Hco-PGP-16 transport activity was studied using the fluorophore Rhodamine 123 (Rho 123). Transfected cells expressing Hco-PGP-16 accumulated less than 50% of Rho 123 than control cells, suggesting an active transport of this tracer dye by Hco-PGP-16. The influence of the MLs on the Rho123 transport by Hco-PGP-16 was then investigated. A marked inhibition of Rho123 transport by ABA and IVM was observed. In contrast, MOX showed less effect on inhibition of Rho123 transport by Hco-PGP-16, and the inhibition was not saturable. The difference in the interaction of the avermectins and MOX with Hco-PGP-16 may help explain the slower rate of development of resistance to MOX compared with the avermectins in H. contortus.