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Mechanisms of Resistance

Role of pfmdr1 Amplification and Expression in Induction of Resistance to Artemisinin Derivatives in Plasmodium falciparum

Marina Chavchich, Lucia Gerena, Jennifer Peters, Nanhua Chen, Qin Cheng, Dennis E. Kyle
Marina Chavchich
1Australian Army Malaria Institute, Weary Dunlop Drive, Gallipoli Barracks, Enoggera, Q4051
2Queensland Institute of Medical Research, PO Royal Brisbane Hospital, Herston, Q4029, Australia
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Lucia Gerena
3Division of Experimental Therapeutics, Walter Reed Army Institute of Research, Silver Spring, Maryland 20910
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Jennifer Peters
1Australian Army Malaria Institute, Weary Dunlop Drive, Gallipoli Barracks, Enoggera, Q4051
2Queensland Institute of Medical Research, PO Royal Brisbane Hospital, Herston, Q4029, Australia
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Nanhua Chen
1Australian Army Malaria Institute, Weary Dunlop Drive, Gallipoli Barracks, Enoggera, Q4051
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Qin Cheng
1Australian Army Malaria Institute, Weary Dunlop Drive, Gallipoli Barracks, Enoggera, Q4051
2Queensland Institute of Medical Research, PO Royal Brisbane Hospital, Herston, Q4029, Australia
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Dennis E. Kyle
1Australian Army Malaria Institute, Weary Dunlop Drive, Gallipoli Barracks, Enoggera, Q4051
3Division of Experimental Therapeutics, Walter Reed Army Institute of Research, Silver Spring, Maryland 20910
4Department of Global Health, College of Public Health, University of South Florida, 3720 Spectrum Blvd., Tampa, Florida 33612
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  • For correspondence: dkyle@health.usf.edu
DOI: 10.1128/AAC.00947-09
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  • FIG. 1.
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    FIG. 1.

    Changes in the index of resistance (ratio of the IC50 for the resistant progeny to the IC50 for the parental line) to various drugs during the adaptation of the Plasmodium falciparum lines W2 (A), TM91C235 (B), and D6 (C) to artelinic acid (AL).

  • FIG. 2.
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    FIG. 2.

    pfmdr1 copy numbers in parental parasites and resistant progenies.

  • FIG. 3.
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    FIG. 3.

    Southern blot hybridization of P. falciparum chromosomes digested with SmaI (left) and BglI (right) and hybridized with probes derived from the pfmdr1 (A) and aldolase (B) genes.

  • FIG. 4.
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    FIG. 4.

    Transcription of the pfmdr1 gene in the W2, W2AL80, W2QHS40, TM91C235, TM91C235AL80, D6, and D6AL80 lines of P. falciparum.

  • FIG. 5.
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    FIG. 5.

    MDR1 protein expression in resistant and parental Plasmodium falciparum lines. Western blot hybridization was performed with anti-Pgh1 (α1435; left side in panels A and B) and anti-RAP2 (6C1; right side in panels A and B) antibodies.

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  • TABLE 1.

    In vitro susceptibilities of Plasmodium falciparum lines to artelinic acid

    Parasite lineMean IC (nM) ± SDa
    50%90%
    W24.73 ± 0.197.45 ± 0.40
    W2AL5023.30 ± 3.9551.31 ± 1.31
    W2AL8022.75 ± 5.3756.80 ± 5.44
    D69.81 ± 1.4121.83 ± 3.04
    D6AL5034.00 ± 3.1481.26 ± 8.67
    D6AL8023.64 ± 3.7175.89 ± 2.60
    TM91C23511.08 ± 1.6822.99 ± 2.60
    TM91C235AL5033.72 ± 1.6453.51 ± 1.12
    TM91C235AL8024.55 ± 4.1375.89 ± 6.8
    • ↵ a n, ≥3.

  • TABLE 2.

    pfmdr1 allelic types in parental and resistant Plasmodium falciparum lines

    Parasite lineAmino acid at position:
    86184103410421246
    D6NYSND
    D6AL80NYSND
    TM91C235NFSND
    TM91C235AL80NFSND
    W2YYSND
    W2AL200YYSND
    W2QHS200YYSND
    W2AS80YYSND

Additional Files

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    Files in this Data Supplement:

    • Supplemental file 1 - Table S1, Primers used for PCR amplification, sequencing, and real-time PCR. Table S2, information on the repetitive region in PfUBCTH.
      Zipped MS Word document, 25K.
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Role of pfmdr1 Amplification and Expression in Induction of Resistance to Artemisinin Derivatives in Plasmodium falciparum
Marina Chavchich, Lucia Gerena, Jennifer Peters, Nanhua Chen, Qin Cheng, Dennis E. Kyle
Antimicrobial Agents and Chemotherapy May 2010, 54 (6) 2455-2464; DOI: 10.1128/AAC.00947-09

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Role of pfmdr1 Amplification and Expression in Induction of Resistance to Artemisinin Derivatives in Plasmodium falciparum
Marina Chavchich, Lucia Gerena, Jennifer Peters, Nanhua Chen, Qin Cheng, Dennis E. Kyle
Antimicrobial Agents and Chemotherapy May 2010, 54 (6) 2455-2464; DOI: 10.1128/AAC.00947-09
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KEYWORDS

antimalarials
artemisinins
drug resistance
Genes, Protozoan
Multidrug Resistance-Associated Proteins
Plasmodium falciparum

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