Toxoplasma gondii: A simple high-throughput assay for drug screening in vitro

ChunMei Jin, Kusuma Kaewintajuk, JingHua Jiang, WooJin Jeong, Masaki Kamata, Hye-Sook Kim, Yusuke Wataya, Hyun Park

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Toxoplasma gondii is the etiologic agent of toxoplasmosis. Although the combination of sulfadiazine and pyrimethamine is used as therapy for this disease, these drugs can have serious side effects and its use is limited in pregnancy. Therefore there is a need for new anti-T. gondii drugs in the clinic. Some systems for T. gondii drug screening have been described, but these have limitations and can be difficult. In order to solve these problems, we established a system to screen drugs in vitro that involved using cell viability methods to calculate drug selectivities, which are Trypan blue, [3-(4,5-dimethylthiazol-zyl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazoliuzolium, inner salt] (MTS) method and lactate dehydrogenase (LDH) assay. These assays were simple to establish and perform. The IC50 values calculated from the morphological assay were not significantly different from the EC50 values calculated using the other three methods. In particular, the results of the morphological assay showed a distinct association with the MTS assay (R = 0.9841). These assays could be used for a wide range of applications in the screening of new drugs and may provide an alternative to the techniques currently used to screen for candidate anti-T. gondii compounds in vitro. In this study, we also tested many compounds and identified some that had a good anti-T. gondii effect in vitro based on the MTS assay. This simple and fast system allowed us to determine which compounds to investigate further using in vivo experiments.

Original languageEnglish
Pages (from-to)132-136
Number of pages5
JournalExperimental Parasitology
Volume121
Issue number2
DOIs
Publication statusPublished - Feb 2009

Fingerprint

Preclinical Drug Evaluations
Toxoplasma
Pharmaceutical Preparations
Sulfadiazine
Pyrimethamine
Trypan Blue
Toxoplasmosis
L-Lactate Dehydrogenase
Inhibitory Concentration 50
Cell Survival
Salts
Drug Therapy
Pregnancy
In Vitro Techniques
Thomsen-Friedenreich antibodies

Keywords

  • LDH assay
  • MTS method
  • Toxoplasma gondii drug screening system
  • Trypan blue dye

ASJC Scopus subject areas

  • Parasitology
  • Immunology

Cite this

Toxoplasma gondii : A simple high-throughput assay for drug screening in vitro. / Jin, ChunMei; Kaewintajuk, Kusuma; Jiang, JingHua; Jeong, WooJin; Kamata, Masaki; Kim, Hye-Sook; Wataya, Yusuke; Park, Hyun.

In: Experimental Parasitology, Vol. 121, No. 2, 02.2009, p. 132-136.

Research output: Contribution to journalArticle

Jin, C, Kaewintajuk, K, Jiang, J, Jeong, W, Kamata, M, Kim, H-S, Wataya, Y & Park, H 2009, 'Toxoplasma gondii: A simple high-throughput assay for drug screening in vitro', Experimental Parasitology, vol. 121, no. 2, pp. 132-136. https://doi.org/10.1016/j.exppara.2008.10.006
Jin, ChunMei ; Kaewintajuk, Kusuma ; Jiang, JingHua ; Jeong, WooJin ; Kamata, Masaki ; Kim, Hye-Sook ; Wataya, Yusuke ; Park, Hyun. / Toxoplasma gondii : A simple high-throughput assay for drug screening in vitro. In: Experimental Parasitology. 2009 ; Vol. 121, No. 2. pp. 132-136.
@article{7b117821cfc04f8dbdc3d3c6f72da315,
title = "Toxoplasma gondii: A simple high-throughput assay for drug screening in vitro",
abstract = "Toxoplasma gondii is the etiologic agent of toxoplasmosis. Although the combination of sulfadiazine and pyrimethamine is used as therapy for this disease, these drugs can have serious side effects and its use is limited in pregnancy. Therefore there is a need for new anti-T. gondii drugs in the clinic. Some systems for T. gondii drug screening have been described, but these have limitations and can be difficult. In order to solve these problems, we established a system to screen drugs in vitro that involved using cell viability methods to calculate drug selectivities, which are Trypan blue, [3-(4,5-dimethylthiazol-zyl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazoliuzolium, inner salt] (MTS) method and lactate dehydrogenase (LDH) assay. These assays were simple to establish and perform. The IC50 values calculated from the morphological assay were not significantly different from the EC50 values calculated using the other three methods. In particular, the results of the morphological assay showed a distinct association with the MTS assay (R = 0.9841). These assays could be used for a wide range of applications in the screening of new drugs and may provide an alternative to the techniques currently used to screen for candidate anti-T. gondii compounds in vitro. In this study, we also tested many compounds and identified some that had a good anti-T. gondii effect in vitro based on the MTS assay. This simple and fast system allowed us to determine which compounds to investigate further using in vivo experiments.",
keywords = "LDH assay, MTS method, Toxoplasma gondii drug screening system, Trypan blue dye",
author = "ChunMei Jin and Kusuma Kaewintajuk and JingHua Jiang and WooJin Jeong and Masaki Kamata and Hye-Sook Kim and Yusuke Wataya and Hyun Park",
year = "2009",
month = "2",
doi = "10.1016/j.exppara.2008.10.006",
language = "English",
volume = "121",
pages = "132--136",
journal = "Experimental Parasitology",
issn = "0014-4894",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Toxoplasma gondii

T2 - A simple high-throughput assay for drug screening in vitro

AU - Jin, ChunMei

AU - Kaewintajuk, Kusuma

AU - Jiang, JingHua

AU - Jeong, WooJin

AU - Kamata, Masaki

AU - Kim, Hye-Sook

AU - Wataya, Yusuke

AU - Park, Hyun

PY - 2009/2

Y1 - 2009/2

N2 - Toxoplasma gondii is the etiologic agent of toxoplasmosis. Although the combination of sulfadiazine and pyrimethamine is used as therapy for this disease, these drugs can have serious side effects and its use is limited in pregnancy. Therefore there is a need for new anti-T. gondii drugs in the clinic. Some systems for T. gondii drug screening have been described, but these have limitations and can be difficult. In order to solve these problems, we established a system to screen drugs in vitro that involved using cell viability methods to calculate drug selectivities, which are Trypan blue, [3-(4,5-dimethylthiazol-zyl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazoliuzolium, inner salt] (MTS) method and lactate dehydrogenase (LDH) assay. These assays were simple to establish and perform. The IC50 values calculated from the morphological assay were not significantly different from the EC50 values calculated using the other three methods. In particular, the results of the morphological assay showed a distinct association with the MTS assay (R = 0.9841). These assays could be used for a wide range of applications in the screening of new drugs and may provide an alternative to the techniques currently used to screen for candidate anti-T. gondii compounds in vitro. In this study, we also tested many compounds and identified some that had a good anti-T. gondii effect in vitro based on the MTS assay. This simple and fast system allowed us to determine which compounds to investigate further using in vivo experiments.

AB - Toxoplasma gondii is the etiologic agent of toxoplasmosis. Although the combination of sulfadiazine and pyrimethamine is used as therapy for this disease, these drugs can have serious side effects and its use is limited in pregnancy. Therefore there is a need for new anti-T. gondii drugs in the clinic. Some systems for T. gondii drug screening have been described, but these have limitations and can be difficult. In order to solve these problems, we established a system to screen drugs in vitro that involved using cell viability methods to calculate drug selectivities, which are Trypan blue, [3-(4,5-dimethylthiazol-zyl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazoliuzolium, inner salt] (MTS) method and lactate dehydrogenase (LDH) assay. These assays were simple to establish and perform. The IC50 values calculated from the morphological assay were not significantly different from the EC50 values calculated using the other three methods. In particular, the results of the morphological assay showed a distinct association with the MTS assay (R = 0.9841). These assays could be used for a wide range of applications in the screening of new drugs and may provide an alternative to the techniques currently used to screen for candidate anti-T. gondii compounds in vitro. In this study, we also tested many compounds and identified some that had a good anti-T. gondii effect in vitro based on the MTS assay. This simple and fast system allowed us to determine which compounds to investigate further using in vivo experiments.

KW - LDH assay

KW - MTS method

KW - Toxoplasma gondii drug screening system

KW - Trypan blue dye

UR - http://www.scopus.com/inward/record.url?scp=57949103535&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=57949103535&partnerID=8YFLogxK

U2 - 10.1016/j.exppara.2008.10.006

DO - 10.1016/j.exppara.2008.10.006

M3 - Article

C2 - 18977350

AN - SCOPUS:57949103535

VL - 121

SP - 132

EP - 136

JO - Experimental Parasitology

JF - Experimental Parasitology

SN - 0014-4894

IS - 2

ER -