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BioAssay: AID 624298

Activators of T cell receptors: Hit Validation

Normally activation of naive T cells requires two signals. 1. A signal through the T cell receptor (TCR) which provides antigen specificity to the response. 2. Signals through co-stimulatory molecules, which are necessary for proliferation, survival and cytokine production. In the absence of co-stimulatory signals T cells are not effectively activated, and may even be rendered anergic or more ..
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 Tested Compounds
 Tested Compounds
All(231)
 
 
Active(13)
 
 
Inactive(161)
 
 
Inconclusive(57)
 
 
 Tested Substances
 Tested Substances
All(231)
 
 
Active(13)
 
 
Inactive(161)
 
 
Inconclusive(57)
 
 
AID: 624298
Data Source: NCGC (Tcell201)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2012-06-15

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 13
Related Experiments
AIDNameTypeComment
504925Activators of T cell receptors: SummarySummarydepositor-specified cross reference: Summary AID
504894Activators of T cell receptors: qHTS campaignConfirmatorysame project related to Summary assay
Description:
Normally activation of naive T cells requires two signals. 1. A signal through the T cell receptor (TCR) which provides antigen specificity to the response. 2. Signals through co-stimulatory molecules, which are necessary for proliferation, survival and cytokine production. In the absence of co-stimulatory signals T cells are not effectively activated, and may even be rendered anergic or unresponsive to subsequent signaling. This lack of costimulation has been shown in multiple model systems to be a major factor in the limited strength and effectiveness of anti-tumor T cell responses. The goal of this project is to identify small molecule compounds that will stimulate T cell activation in the presence of TCR signaling but in the absence of co-stimulatory signals, thereby allowing activation of naive T cells in the absence of co-stimulation. Identification of such compounds would have clear experimental and clinical applications in instances where desirable antigen-specific immune responses are not generated due to a lack of co-stimulation, for example in the generation of anti-tumor responses.

In collaboration between the National Cancer Institute and NIH Chemical Genomics Center, a high-throughput amenable screen was developed to discover small molecules that co-stimulate and activate human T cell receptors. The screen will be carried out using a novel homogeneous time resolved fluorescence energy transfer (HTRF) assay to measure production of the cytokine IL-2 by activated T cells and will be tested against the NIH Molecular Libraries Small Molecule Repository (MLSMR). This is the same assay tested against the hits identified in the primary screen.

NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Probe Centers Network [MLPCN]

MLPCN Grant: MH090820
Assay Submitter (PI): Richard Hodes, National Cancer Institute, NIH
Protocol
Primary human leukocytes are incubated with a-CD3 beads and compounds for 48 hr. The amount of secreted IL-2 is then assayed with a HTRF IL-2 kit. The presence of IL-2 cytokine captures the detection reagents and brings the a-IL-2-K and a-IL-2-d2 into close proximity. Upon excitation with 320nm light, the europium cryptate is capable of transferring energy to acceptor molecules within close proximity, resulting in acceptor molecule emission at 665nm.
Comment
Compound Ranking:

1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent activators are ranked higher than compounds that showed apparent inhibition.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.
Categorized Comment - additional comments and annotations
From PubChem:
Assay Cell Type: T cell
From ChEMBL:
Assay Type: Functional
Assay Cell Type: NULL
Result Definitions
Show more
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1PhenotypeIndicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
2Potency*Concentration at which compound exhibits half-maximal efficacy, AC50. Extrapolated AC50s also include the highest efficacy observed and the concentration of compound at which it was observed.FloatμM
3EfficacyMaximal efficacy of compound, reported as a percentage of control. These values are estimated based on fits of the Hill equation to the dose-response curves.Float%
4Analysis CommentAnnotation/notes on a particular compound's data or its analysis.String
5Activity_ScoreActivity score.Integer
6Curve_DescriptionA description of dose-response curve quality. A complete curve has two observed asymptotes; a partial curve may not have attained its second asymptote at the highest concentration tested. High efficacy curves exhibit efficacy greater than 80% of control. Partial efficacies are statistically significant, but below 80% of control.String
7Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
8Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
9Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
10Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
11Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
12Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
13Excluded_PointsWhich dose-response titration points were excluded from analysis based on outlier analysis. Each number represents whether a titration point was (1) or was not (0) excluded, for the titration series going from smallest to highest compound concentrations.String
14Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
15Activity at 0.0002601382 uM (0.000260138μM**)% Activity at given concentration.Float%
16Activity at 0.0007804147 uM (0.000780415μM**)% Activity at given concentration.Float%
17Activity at 0.00234 uM (0.00234124μM**)% Activity at given concentration.Float%
18Activity at 0.00702 uM (0.00702378μM**)% Activity at given concentration.Float%
19Activity at 0.021 uM (0.0210713μM**)% Activity at given concentration.Float%
20Activity at 0.063 uM (0.0630298μM**)% Activity at given concentration.Float%
21Activity at 0.090 uM (0.0900058μM**)% Activity at given concentration.Float%
22Activity at 0.190 uM (0.189557μM**)% Activity at given concentration.Float%
23Activity at 0.360 uM (0.360023μM**)% Activity at given concentration.Float%
24Activity at 0.570 uM (0.570077μM**)% Activity at given concentration.Float%
25Activity at 1.440 uM (1.44009μM**)% Activity at given concentration.Float%
26Activity at 1.707 uM (1.70678μM**)% Activity at given concentration.Float%
27Activity at 2.880 uM (2.88018μM**)% Activity at given concentration.Float%
28Activity at 5.126 uM (5.12551μM**)% Activity at given concentration.Float%
29Activity at 11.52 uM (11.5207μM**)% Activity at given concentration.Float%
30Activity at 15.36 uM (15.361μM**)% Activity at given concentration.Float%
31Activity at 23.04 uM (23.0415μM**)% Activity at given concentration.Float%
32Activity at 46.08 uM (46.0829μM**)% Activity at given concentration.Float%
33Activity at 92.17 uM (92.1659μM**)% Activity at given concentration.Float%
34Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: MH090820

Data Table (Concise)
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