Bookmark and Share
BioAssay: AID 652279

Cidal vs. static assay: inhibition of T. cruzi (CAI/72) in J774 macrophages after prolonged compound treatment and treatment withdrawal Measured in Cell-Based System Using Imaging - 2138-13_Inhibitor_SinglePoint_DryPowder_Activity

Assay Overview: This assay was conducted to determine if the compounds are cidal to Trypanosoma cruzi cells (amastigotes) within the mammalian J774 macrophages. After 24 hours, cells were washed and compounds were added fresh every 3 days for 1 week (protocol 1) or for 3 weeks (protocol 2). After compound treatment, compound was removed from the infected cells and observed for 31 days (protocol more ..
_
   
 Tested Compounds
 Tested Compounds
All(11)
 
 
Active(11)
 
 
 Tested Substances
 Tested Substances
All(11)
 
 
Active(11)
 
 
AID: 652279
Data Source: Broad Institute (2138-13_Inhibitor_SinglePoint_DryPowder_Activity)
Depositor Category: NIH Molecular Libraries Screening Center Network, Assay Provider
BioAssay Version:
Deposit Date: 2013-04-04
Hold-until Date: 2013-07-17
Modify Date: 2013-07-17

Data Table ( Complete ):           View Active Data    View All Data
BioActive Compounds: 11
Related Experiments
Show more
AIDNameTypeProbeComment
624280Broad Institute Inhibition of T.cruzi replication in culture Inhibitor Probe ProjectSummary1 depositor-specified cross reference: Summary
624255Inhibition of T.cruzi proliferation in culture Measured in Cell-Based System Using Plate Reader - 2138-01_Inhibitor_SinglePoint_HTS_ActivityScreening same project related to Summary assay
651739Inhibition of T.cruzi proliferation in culture Measured in Cell-Based System Using Plate Reader - 2138-01_Inhibitor_SinglePoint_CherryPick_ActivityOther same project related to Summary assay
651740Inhibition of T.cruzi proliferation in culture Measured in Cell-Based System Using Plate Reader - 2138-01_Inhibitor_SinglePoint_CherryPick_Activity_Set2Other same project related to Summary assay
651742NIH/3T3 (mouse embryonic fibroblast) toxicity Measured in Cell-Based System Using Plate Reader - 2138-02_Inhibitor_SinglePoint_CherryPick_ActivityOther same project related to Summary assay
651744NIH/3T3 (mouse embryonic fibroblast) toxicity Measured in Cell-Based System Using Plate Reader - 2138-02_Inhibitor_SinglePoint_CherryPick_Activity_Set2Other same project related to Summary assay
651817NIH/3T3 (mouse embryonic fibroblast ) toxicity Measured in Cell-Based System Using Plate Reader - 2017-02_Inhibitor_Dose_CherryPick_Activity_Set2Confirmatory same project related to Summary assay
651818Intracellular Trypanosomes Measured in Cell-Based/Microorganism System Using Plate Reader - 2017-01_Inhibitor_Dose_CherryPick_Activity_Set3Confirmatory same project related to Summary assay
651844NIH/3T3 (mouse embryonic fibroblast) toxicity Measured in Cell-Based System Using Plate Reader - 2138-02_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651845Inhibition of T.cruzi proliferation in culture Measured in Cell-Based System Using Plate Reader - 2138-01_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651869Cidal vs. static assay: inhibition of T. cruzi (CAI/72) after prolonged treatment and treatment withdrawal Measured in Cell-Based System Using Imaging - 2138-08_Inhibitor_SinglePoint_DryPowder_Activity_Set2Other same project related to Summary assay
651877Cidal vs. static assay: inhibition of T. cruzi (CAI/72) after prolonged treatment and treatment withdrawal Measured in Cell-Based System Using Imaging - 2138-08_Inhibitor_SinglePoint_DryPowder_ActivityOther same project related to Summary assay
651881Inhibition of T. cruzi Tulahuen strain amastigotes in L-6 muscle cells Measured in Cell-Based and Microorganism Combination System Using Plate Reader - 2138-03_Inhibitor_Dose_CherryPick_ActivityConfirmatory same project related to Summary assay
651885Inhibition of intracellular CAI/72 strain of Trypanosoma cruzi in bovine embryo skeletal muscle cells: high content imaging Measured in Cell-Based System Using Imaging - 2138-07_Inhibitor_SinglePoint_DryPowder_ActivityConfirmatory same project related to Summary assay
651888Inhibition of Plasmodium falcipirum NF54 Measured in Cell-Based System Using Plate Reader - 2138-11_Inhibitor_Dose_CherryPick_ActivityConfirmatory same project related to Summary assay
651889Inhibition of Plasmodium falcipirum NF54 Measured in Cell-Based System Using Plate Reader - 2138-11_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651890Inhibition of T. cruzi Tulahuen strain amastigotes in L-6 muscle cells Measured in Cell-Based and Microorganism Combination System Using Plate Reader - 2138-03_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651891Inhibition of L. donovani axenic amastigotes Measured in Cell-Based System Using Plate Reader - 2138-04_Inhibitor_Dose_CherryPick_ActivityConfirmatory same project related to Summary assay
651892Inhibition of L. donovani axenic amastigotes Measured in Cell-Based System Using Plate Reader - 2138-04_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651893Inhibition of Trypanosoma brucei rhodesiense STIB900 Measured in Cell-Based System Using Plate Reader - 2138-05_Inhibitor_Dose_CherryPick_ActivityConfirmatory same project related to Summary assay
651894Inhibition of Trypanosoma brucei rhodesiense STIB900 Measured in Cell-Based System Using Plate Reader - 2138-05_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651895L-6 rat myocyte cell toxicity Measured in Cell-Based System Using Plate Reader - 2138-06_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651896L-6 rat myocyte cell toxicity Measured in Cell-Based System Using Plate Reader - 2138-06_Inhibitor_Dose_CherryPick_ActivityConfirmatory same project related to Summary assay
651897Inhibition of Trypanosoma cruzi Cruzain Measured in Biochemical System Using Plate Reader - 2138-09_Inhibitor_SinglePoint_DryPowder_ActivityConfirmatory same project related to Summary assay
651903Intracellular Trypanosomes Measured in Cell-Based/Microorganism System Using Plate Reader - 2017-01_Inhibitor_SinglePoint_HTS_Activity_Set2Screening same project related to Summary assay
652275Inhibition of T.cruzi proliferation in culture Measured in Cell-Based System Using Plate Reader - 2138-01_Inhibitor_Dose_DryPowder_Activity_Set2Confirmatory same project related to Summary assay
652278Cidal vs. static assay: inhibition of T. cruzi (CAI/72) in J774 macrophages after prolonged compound treatment and treatment withdrawal Measured in Cell-Based System Using Imaging - 2138-13_Inhibitor_SinglePoint_DryPowder_Activity_Set2Other same project related to Summary assay
652280NIH/3T3 (mouse embryonic fibroblast) toxicity Measured in Cell-Based System Using Plate Reader - 2138-02_Inhibitor_Dose_DryPowder_Activity_Set2Confirmatory same project related to Summary assay
686919Inhibition of Trypanosoma cruzi Cruzain Measured in Biochemical System Using Plate Reader - 2138-09_Inhibitor_Dose_DryPowder_ActivityOther same project related to Summary assay
Description:
Keywords: Trypanosoma cruzi, amastigotes, cidal


Assay Overview: This assay was conducted to determine if the compounds are cidal to Trypanosoma cruzi cells (amastigotes) within the mammalian J774 macrophages. After 24 hours, cells were washed and compounds were added fresh every 3 days for 1 week (protocol 1) or for 3 weeks (protocol 2). After compound treatment, compound was removed from the infected cells and observed for 31 days (protocol 1) or 17 days (protocol 2). The emergence of fully developed trypomastigotes was assessed daily via light microscope. The mammalian T.cruzi life cycle is 6 days; when treated with DMSO or a nonactive compound, 6 days after infection is the earliest possible emergence of trypomastigotes. If a compound lyses all intracellular amastigotes, no trypomastigotes will emerge at any day during compound treatment or compound withdrawal. However, if a compound is static or only partially lytic, there will be a delay of trypomastigote emergence beyond the traditional 6 days. For graphic representation, day "0" is the first day of compound withdrawl. For protocol 1, compound treatment starts on day '-7' and for protocol 2, compound treatment starts on day '-21'. Therefore the trypomastigotes appear at day '-1' for protocol 1 and day '-15' for protocol 2 when treated with DMSO only.


Expected Outcome: For those compounds that are cidal, no trypomastigotes will appear at any day tested with or without compound treatment. For those compounds that have a static effect, it is expected that compound treatment will suppress replication during treatment, but upon compound removal, trypomastigotes will immediately reappear. Compounds that alter the normal replication of T. cruzi may extend the normal life cycle past 6 days even after compound removal.
Protocol
This assay assesses cidal activity of compounds in T. cruzi (CAI/72 )-infected irradiated J774 macrophages . J774 macrophages cultured in RPMI-1640 medium with 5% heat inactivated fetal calf serum (FCS) were irradiated (9000 rad) to arrest the cell cycle and plated onto 48-well tissue culture plates and then infected with the CAI/72 strain of Trypanosoma cruzi. Twenty four hours after infection, infected cells are washed and compounds are added every three days for 1 week (protocol 1) or 3 weeks (protocol 2). After the initial period, compounds are removed and cells are observed for an additional 31 days (protocol 1) or 17 days (protocol 2) without drug. During which time cultures are monitored for reappearance of T. cruzi parasites. This procedure allows us to determine if host cells were effectively cured (i.e., compound is cidal) or if infection still persists (i.e, static activity).

To analyze the data, the day of trypomastigote reemergence was added to a linear scale between -7 (1st day of compound treatment) to 31 (final day of experiment) for protocol 1 or -21 (1st day of compound addition) to 17 (final day of experiment) for protocol 2. Concentrations vs. day of emergence were plotted in Graphpad prism and a linear regression was conducted to determine the slope. At the first concentration where no trypomastigotes appeared, the last day of the experiment was added (i.e., 31 for protocol 1 or 17for protocol 2).
Comment
Pubchem Activity Score was calculated by dividing the slope of the test compound by the slope of Benznidazole . Larger numbers are more active and lower numbers are less active.
Categorized Comment - additional comments and annotations
From PubChem:
Assay Format: Cell-based
Assay Cell Type: J774
Result Definitions
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1SlopeSlope of linear regression of compound concentrations vs day of trypomastigote emergence.Float
Additional Information
Grant Number: 1 R03 MH085673-01

Data Table (Concise)
Data Table ( Complete ):     View Active Data    View All Data
PageFrom: