Reliability of Antimalarial Sensitivity Tests Depends on Drug Mechanisms of Action

Overview

Journal J Clin Microbiol

Specialty Microbiology

Date 2010 Mar 12

PMID 20220159

Citations 39

Authors

Sharon Wein

Marjorie Maynadier

Christophe Tran Van Ba

Rachel Cerdan

Suzanne Peyrottes

Laurent Fraisse

Henri Vial

Affiliations

Soon will be listed here.

Abstract

In vitro antimalarial activity tests play a pivotal role in malaria drug research or for monitoring drug resistance in field isolates. We applied two isotopic tests, two enzyme-linked immunosorbent assays (ELISA) and the SYBR green I fluorescence-based assay, to test artesunate and chloroquine, the metabolic inhibitors atovaquone and pyrimethamine, our fast-acting choline analog T3/SAR97276, and doxycycline, which has a delayed death profile. Isotopic tests based on hypoxanthine and ethanolamine incorporation are the most reliable tests provided when they are applied after one full 48-h parasite cycle. The SYBR green assay, which measures the DNA content, usually requires 72 h of incubation to obtain reliable results. When delayed death is suspected, specific protocols are required with increasing incubation times up to 96 h. In contrast, both ELISA tests used (pLDH and HRP2) appear to be problematic, leading to disappointing and even erroneous results for molecules that do not share an artesunatelike profile. The reliability of these tests is linked to the mode of action of the drug, and the conditions required to get informative results are hard to predict. Our results suggest some minimal conditions to apply these tests that should give rise to a standard 50% inhibitory concentration, regardless of the mechanism of action of the compounds, and highlight that the most commonly used in vitro antimalarial activity tests do not have the same potential. Some of them might not detect the antimalarial potential of new classes of compounds with innovative modes of action, which subsequently could become promising new antimalarial drugs.

Citing Articles

Plant Extracts as a Source of Natural Products with Potential Antimalarial Effects: An Update from 2018 to 2022.

Ribeiro G, Rei Yan S, Palmisano G, Wrenger C Pharmaceutics. 2023; 15(6).

PMID: 37376086 PMC: 10300920. DOI: 10.3390/pharmaceutics15061638.

Rapid and quantitative antimalarial drug efficacy testing via the magneto-optical detection of hemozoin.

Molnar P, Orban A, Izrael R, Babai R, Marton L, Butykai A Sci Rep. 2020; 10(1):14025.

PMID: 32820190 PMC: 7441145. DOI: 10.1038/s41598-020-70860-y.

Real-time cholesterol sorting in Plasmodium falciparum-erythrocytes as revealed by 3D label-free imaging.

Hayakawa E, Yamaguchi K, Mori M, Nardone G Sci Rep. 2020; 10(1):2794.

PMID: 32066816 PMC: 7026401. DOI: 10.1038/s41598-020-59552-9.

Antiplasmodial natural products: an update.

Tajuddeen N, Van Heerden F Malar J. 2019; 18(1):404.

PMID: 31805944 PMC: 6896759. DOI: 10.1186/s12936-019-3026-1.

Low polymorphisms in pfact, pfugt and pfcarl genes in African Plasmodium falciparum isolates and absence of association with susceptibility to common anti-malarial drugs.

Foguim F, Robert M, Gueye M, Gendrot M, Diawara S, Mosnier J Malar J. 2019; 18(1):293.

PMID: 31455301 PMC: 6712813. DOI: 10.1186/s12936-019-2919-3.

References

Smilkstein M, Sriwilaijaroen N, Kelly J, Wilairat P, Riscoe M . Simple and inexpensive fluorescence-based technique for high-throughput antimalarial drug screening. Antimicrob Agents Chemother. 2004; 48(5):1803-6. PMC: 400546. DOI: 10.1128/AAC.48.5.1803-1806.2004. View

Ancelin M, Calas M, Vidal-Sailhan V, Herbute S, Ringwald P, Vial H . Potent inhibitors of Plasmodium phospholipid metabolism with a broad spectrum of in vitro antimalarial activities. Antimicrob Agents Chemother. 2003; 47(8):2590-7. PMC: 166094. DOI: 10.1128/AAC.47.8.2590-2597.2003. View

Vial H, Wein S, Farenc C, Kocken C, Nicolas O, Ancelin M . Prodrugs of bisthiazolium salts are orally potent antimalarials. Proc Natl Acad Sci U S A. 2004; 101(43):15458-63. PMC: 523447. DOI: 10.1073/pnas.0404037101. View

TRAGER W, Jensen J . Human malaria parasites in continuous culture. Science. 1976; 193(4254):673-5. DOI: 10.1126/science.781840. View

Lambros C, Vanderberg J . Synchronization of Plasmodium falciparum erythrocytic stages in culture. J Parasitol. 1979; 65(3):418-20. View

Desjardins R, Canfield C, Haynes J, Chulay J . Quantitative assessment of antimalarial activity in vitro by a semiautomated microdilution technique. Antimicrob Agents Chemother. 1979; 16(6):710-8. PMC: 352941. DOI: 10.1128/AAC.16.6.710. View

Divo A, Geary T, Jensen J . Oxygen- and time-dependent effects of antibiotics and selected mitochondrial inhibitors on Plasmodium falciparum in culture. Antimicrob Agents Chemother. 1985; 27(1):21-7. PMC: 176198. DOI: 10.1128/AAC.27.1.21. View

Wellems T, Howard R . Homologous genes encode two distinct histidine-rich proteins in a cloned isolate of Plasmodium falciparum. Proc Natl Acad Sci U S A. 1986; 83(16):6065-9. PMC: 386439. DOI: 10.1073/pnas.83.16.6065. View

Carlson J, Helmby H, Hill A, Brewster D, Greenwood B, Wahlgren M . Human cerebral malaria: association with erythrocyte rosetting and lack of anti-rosetting antibodies. Lancet. 1990; 336(8729):1457-60. DOI: 10.1016/0140-6736(90)93174-n. View

10.

Elabbadi N, Ancelin M, Vial H . Use of radioactive ethanolamine incorporation into phospholipids to assess in vitro antimalarial activity by the semiautomated microdilution technique. Antimicrob Agents Chemother. 1992; 36(1):50-5. PMC: 189224. DOI: 10.1128/AAC.36.1.50. View

11.

Vial H, Ancelin M . Malarial lipids. An overview. Subcell Biochem. 1992; 18:259-306. View

12.

Ter Kuile F, White N, Holloway P, Pasvol G, Krishna S . Plasmodium falciparum: in vitro studies of the pharmacodynamic properties of drugs used for the treatment of severe malaria. Exp Parasitol. 1993; 76(1):85-95. DOI: 10.1006/expr.1993.1010. View

13.

Makler M, Ries J, Williams J, Bancroft J, Piper R, Gibbins B . Parasite lactate dehydrogenase as an assay for Plasmodium falciparum drug sensitivity. Am J Trop Med Hyg. 1993; 48(6):739-41. DOI: 10.4269/ajtmh.1993.48.739. View

14.

Skinner T, Manning L, Johnston W, Davis T . In vitro stage-specific sensitivity of Plasmodium falciparum to quinine and artemisinin drugs. Int J Parasitol. 1996; 26(5):519-25. DOI: 10.1016/0020-7519(96)89380-5. View

15.

Ancelin M, Calas M, Bompart J, Cordina G, Martin D, Ben Bari M . Antimalarial activity of 77 phospholipid polar head analogs: close correlation between inhibition of phospholipid metabolism and in vitro Plasmodium falciparum growth. Blood. 1998; 91(4):1426-37. View

16.

Lang-Unnasch N, Murphy A . Metabolic changes of the malaria parasite during the transition from the human to the mosquito host. Annu Rev Microbiol. 1999; 52:561-90. DOI: 10.1146/annurev.micro.52.1.561. View

17.

Biagini G, Pasini E, Hughes R, de Koning H, Vial H, ONeill P . Characterization of the choline carrier of Plasmodium falciparum: a route for the selective delivery of novel antimalarial drugs. Blood. 2004; 104(10):3372-7. DOI: 10.1182/blood-2004-03-1084. View

18.

Noedl H, Bronnert J, Yingyuen K, Attlmayr B, Kollaritsch H, Fukuda M . Simple histidine-rich protein 2 double-site sandwich enzyme-linked immunosorbent assay for use in malaria drug sensitivity testing. Antimicrob Agents Chemother. 2005; 49(8):3575-7. PMC: 1196223. DOI: 10.1128/AAC.49.8.3575-3577.2005. View

19.

Noedl H, Krudsood S, Leowattana W, Tangpukdee N, Thanachartwet W, Looareesuwan S . In vitro antimalarial activity of azithromycin, artesunate, and quinine in combination and correlation with clinical outcome. Antimicrob Agents Chemother. 2006; 51(2):651-6. PMC: 1797729. DOI: 10.1128/AAC.01023-06. View

20.

Sidhu A, Sun Q, Nkrumah L, Dunne M, Sacchettini J, Fidock D . In vitro efficacy, resistance selection, and structural modeling studies implicate the malarial parasite apicoplast as the target of azithromycin. J Biol Chem. 2006; 282(4):2494-504. DOI: 10.1074/jbc.M608615200. View