Toxoplasma gondii: a simple high-throughput assay for drug screening in vitro.
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 IC(50) values calculated from the morphological assay were not significantly different from the EC(50) 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.
Jin
ChunMei
C
Department of Infection Biology, Zoonosis Research Center, Wonkwang University School of Medicine, Chonbuk, Republic of Korea.
Kaewintajuk
Kusuma
K
Jiang
JingHua
J
Jeong
WooJin
W
Kamata
Masaki
M
Kim
Hye-Sook
HS
Wataya
Yusuke
Y
Park
Hyun
H
D000818:Animals; D003049:Coccidiostats / Q000494:pharmacology*; D004396:Coloring Agents; D004353:Drug Evaluation, Preclinical / Q000379:methods* / Q000592:standards; D004797:Enzyme-Linked Immunosorbent Assay; D006367:HeLa Cells; D006801:Humans; D020128:Inhibitory Concentration 50; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D011739:Pyrimethamine / Q000494:pharmacology; D015572:Spiramycin / Q000494:pharmacology; D013411:Sulfadiazine / Q000494:pharmacology; D014122:Toxoplasma / Q000187:drug effects* / Q000502:physiology; D014343:Trypan Blue
D003049:Coccidiostats; D004396:Coloring Agents; D013411:Sulfadiazine; D015572:Spiramycin; D007770:L-Lactate Dehydrogenase; D014343:Trypan Blue; D011739:Pyrimethamine