scholarly journals Visualizing pyrazinamide action by live single cell imaging of phagosome acidification and Mycobacterium tuberculosis pH homeostasis

2021 ◽  
Author(s):  
Pierre Santucci ◽  
Beren Aylan ◽  
Laure Botella ◽  
Elliott M Bernard ◽  
Claudio Bussi ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Cell Imaging ◽  
Intracellular Acidification ◽  
Ph Homeostasis ◽  
Biophysical Properties ◽  
Single Cell Imaging ◽  
Imaging Approach ◽  
Genetic Perturbations ◽  
Dual Imaging ◽  
Antibiotic Efficacy

Mycobacterium tuberculosis (Mtb) segregates within multiple subcellular niches with different biochemical and biophysical properties that, upon treatment, may impact antibiotic distribution, accumulation, and efficacy. However, it remains unclear whether fluctuating intracellular microenvironments alter mycobacterial homeostasis and contribute to antibiotic enrichment and efficacy. Here, we describe a live dual-imaging approach to monitor host subcellular acidification and Mtb intrabacterial pH. By combining this approach with pharmacological and genetic perturbations, we show that Mtb can maintain its intracellular pH independently of the surrounding pH in human macrophages. Importantly, unlike bedaquiline (BDQ), isoniazid (INH) or rifampicin (RIF), the drug pyrazinamide (PZA) displays antibacterial efficacy by acting as protonophore which disrupts intrabacterial pH homeostasis in cellulo. By using Mtb mutants, we confirmed that intracellular acidification is a prerequisite for PZA efficacy in cellulo. We anticipate this imaging approach will be useful to identify host cellular environments that affect antibiotic efficacy against intracellular pathogens.

Single Cell Imaging of Human Cologenic Carcinoma Cells by 3-Tesla MRI

2004 ◽  
Vol 176 (03) ◽  
Author(s):  
UKM Teichgräber ◽  
JG Pinkernelle ◽  
F Neumann ◽  
T Benter ◽  
H Bruhn ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Imaging ◽  
Carcinoma Cells ◽  
3 Tesla ◽  
Single Cell Imaging ◽  
3 Tesla Mri

scholarly journals A high-throughput screening assay based on automated microscopy for monitoring antibiotic susceptibility of Mycobacterium tuberculosis phenotypes

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Sadaf Kalsum ◽  
Blanka Andersson ◽  
Jyotirmoy Das ◽  
Thomas Schön ◽  
Maria Lerm
Keyword(s):  
Mycobacterium Tuberculosis ◽  
High Throughput ◽  
High Throughput Screening ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Imaging System ◽  
Aggregate Size ◽  
Live Cell ◽  
Screening Assay ◽  
High Throughput Screening Assay

Abstract Background Efficient high-throughput drug screening assays are necessary to enable the discovery of new anti-mycobacterial drugs. The purpose of our work was to develop and validate an assay based on live-cell imaging which can monitor the growth of two distinct phenotypes of Mycobacterium tuberculosis and to test their susceptibility to commonly used TB drugs. Results Both planktonic and cording phenotypes were successfully monitored as fluorescent objects using the live-cell imaging system IncuCyte S3, allowing collection of data describing distinct characteristics of aggregate size and growth. The quantification of changes in total area of aggregates was used to define IC50 and MIC values of selected TB drugs which revealed that the cording phenotype grew more rapidly and displayed a higher susceptibility to rifampicin. In checkerboard approach, testing pair-wise combinations of sub-inhibitory concentrations of drugs, rifampicin, linezolid and pretomanid demonstrated superior growth inhibition of cording phenotype. Conclusions Our results emphasize the efficiency of using automated live-cell imaging and its potential in high-throughput whole-cell screening to evaluate existing and search for novel antimycobacterial drugs.

Enabling drug discovery and development through single-cell imaging

2018 ◽  
Vol 14 (2) ◽  
pp. 115-125 ◽  
Author(s):  
Andrea K. Pomerantz ◽  
Farid Sari-Sarraf ◽  
Kerri J. Grove ◽  
Liliana Pedro ◽  
Patrick J. Rudewicz ◽  
...  
Keyword(s):  
Drug Discovery ◽  
Single Cell ◽  
Cell Imaging ◽  
Drug Discovery And Development ◽  
Single Cell Imaging

High-content screening of drug-induced cardiotoxicity using quantitative single cell imaging cytometry on microfluidic device

Lab on a Chip ◽  
2011 ◽  
Vol 11 (1) ◽  
pp. 104-114 ◽  
Author(s):  
Min Jung Kim ◽  
Su Chul Lee ◽  
Sukdeb Pal ◽  
Eunyoung Han ◽  
Joon Myong Song
Keyword(s):  
Single Cell ◽  
Microfluidic Device ◽  
Cell Imaging ◽  
High Content Screening ◽  
Drug Induced ◽  
Single Cell Imaging ◽  
Imaging Cytometry

scholarly journals AE anion exchangers in atrial tumor cells

2001 ◽  
Vol 280 (3) ◽  
pp. H937-H945 ◽  
Author(s):  
Panos Papageorgiou ◽  
Boris E. Shmukler ◽  
Alan K. Stuart-Tilley ◽  
Lianwei Jiang ◽  
Seth L. Alper
Keyword(s):  
Tumor Cell Line ◽  
Disulfonic Acid ◽  
Cell Type ◽  
Intracellular Acidification ◽  
Ph Homeostasis ◽  
Rt Pcr ◽  
Anion Exchangers ◽  
Atrial Tumor ◽  
Atrial Cell ◽  
Exchange Activity

Intracellular pH homeostasis and intracellular Cl−concentration in cardiac myocytes are regulated by anion exchange mechanisms. In physiological extracellular Cl−concentrations, Cl−/HCO[Formula: see text] exchange promotes intracellular acidification and Cl−loading sensitive to inhibition by stilbene disulfonates. We investigated the expression of AE anion exchangers in the AT-1 mouse atrial tumor cell line. Cultured AT-1 cells exhibited a substantial basal Na+-independent Cl−/HCO[Formula: see text] (but not Cl−/OH−) exchange activity that was inhibited by DIDS but not by dibenzamidostilbene disulfonic acid (DBDS). AT-1 cell Cl−/HCO[Formula: see text] activity was stimulated two- to threefold by extracellular ATP and ANG II. AE mRNAs detected by RT-PCR in AT-1 cells included brain AE3 (bAE3), cardiac AE3 (cAE3), AE2a, AE2b, AE2c1, AE2c2, and erythroid AE1 (eAE1), but not kidney AE1 (kAE1). Cultured AT-1 cells expressed AE2, cAE3, and bAE3 polypeptides, which were detected by immunoblot and immunocytochemistry. An AE1-like epitope was detected by immunocytochemistry but not by immunoblot. Both bAE3 and cAE3 were present in intact AT-1 tumors. Cultured AT-1 cells provide a useful system for the study of mediators and regulators of Cl−/HCO[Formula: see text] exchange activity in an atrial cell type.

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