Potential use of nitrate reductase as a biomarker for the identification of active and dormant inhibitors of Mycobacterium tuberculosis in a THP1 infection model

Journal of Biomolecular Screening
Sampa Sarkar, Dhiman Sarkar

Abstract

The development of a macrophage-based, antitubercular high-throughput screening system could expedite discovery programs for identifying novel inhibitors. In this study, the kinetics of nitrate reduction (NR) by Mycobacterium tuberculosis during growth in Thp1 macrophages was found to be almost parallel to viable bacilli count. NR in the culture medium containing 50 mM of nitrate was found to be optimum on the fifth day after infection with M. tuberculosis. The signal-to-noise (S/N) ratio and Z-factor obtained from this macrophage-based assay were 5.4 and 0.965, respectively, which confirms the robustness of the assay protocol. The protocol was further validated by using standard antitubercular inhibitors such as rifampicin, isoniazid, streptomycin, ethambutol, and pyrazinamide, added at their IC(90) value, on the day of infection. These inhibitors were not able to kill the bacilli when added to the culture on the fifth day after infection. Interestingly, pentachlorophenol and rifampicin killed the bacilli immediately after addition on the fifth day of infection. Altogether, this assay protocol using M. tuberculosis-infected Thp-1 macrophages provides a novel, cost-efficient, robust, and easy-to-perform screening platform for t...Continue Reading

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Citations

Mar 2, 2016·Bioorganic & Medicinal Chemistry Letters·Dnyaneshwar D SubhedarBapurao B Shingate
Aug 25, 2015·Bioorganic & Medicinal Chemistry Letters·Vandana S PoreDhiman Sarkar
Dec 20, 2014·The Journal of Antimicrobial Chemotherapy·Vincent DubéeMichel Arthur
Jan 20, 2017·European Journal of Medicinal Chemistry·Mahesh M PisalHanumant B Borate
Oct 17, 2018·Future Medicinal Chemistry·Hemlata M MaliDharmarajan Sriram
Nov 8, 2013·Molecular Based Mathematical Biology·Chuan LiEmil Alexov
Aug 2, 2018·Tropical Medicine and Health·Dristi HalwaiShyamal Kumar Bhattacharya
Jun 3, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Anthi PetrouAthina Geronikaki

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Methods Mentioned

BETA
PMA
Assay

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