{"Id":"7ca8195e-72fc-4923-9168-b6445de3b834","CanonicalId":"","Url":"https://pubmed.ncbi.nlm.nih.gov/32328071/","Name":"Multiplex T Cell stimulation assay utilizing a T Cell activation reporter-based detection system","Description":"This assay can efficiently test antigen reactivity of multiple T cell receptors (TCRs) with limited cost, time, and labor. Nuclear factor of activated T cells (NFAT) is a transcription factor involved in producing cytokines and is often utilized as a reporter system for T cell activation. Using a NFAT-based fluorescent reporter system, we generated T-hybridoma cell lines that express intensely fluorescent proteins in response to antigen stimulation and constitutively express additional fluorescent proteins, which serve as identifiers of each T-hybridoma expressing a unique TCR. This allows for the combination of multiple T-hybridoma lines within a single reaction. Sensitivity to stimulation is not decreased by adding fluorescent proteins or multiplexing T cells. In multiplexed reactions, response by one cell line does not induce response in others, thus preserving specificity. This multiplex assay system will be a useful tool for antigen discovery research in a variety of contexts, including using combinatorial peptide libraries to determine T cell epitopes. ","ResourceType":"AssayTechnology","ParentResourceType":"Technology","DefiningManuscriptId":"PMID32328071","Publications":[],"Contributors":[{"Role":{"CanonicalId":"hirn-cv-ResourceRole-producer","Url":"resources.hirnetwork.org/api/v1/CV/","Name":"Producer","OntologyId":"HIRN"},"HirnUserId":"aaron_michels@hirnetwork.org","FirstName":"Aaron","LastName":"Michels","EmailAddress":"aaron.michels@ucdenver.edu","Organization":null,"Consortia":["CMAI"]},{"Role":{"CanonicalId":"hirn-cv-ResourceRole-producer","Url":"resources.hirnetwork.org/api/v1/CV/","Name":"Producer","OntologyId":"HIRN"},"HirnUserId":"maki_nakayama@hirnetwork.org","FirstName":"Maki","LastName":"Nakayama","EmailAddress":"maki.nakayama@ucdenver.edu","Organization":null,"Consortia":["CMAI"]}],"Vendor":{"id":null,"Name":"","Url":""},"VendorResourceName":"","VendorResourceUrl":"","VendorCatNumber":"","VendorLotNumber":"","DateCreated":"2021-04-30T01:28:29.6140283+00:00","DateModified":"2022-07-01T03:35:02.3718439+00:00","VersionId":"1","ModelVersion":"4.0","TechnologyData":{"Dependencies":[],"AssayTechnologyData":{"BioSampleType":[],"CellsTestedWith":[],"AssayTarget":{"CanonicalId":"","Url":"","Name":"","OntologyId":""},"AssayCategory":{"CanonicalId":"","Url":"","Name":"","OntologyId":""},"TechnologyType":{"CanonicalId":"","Url":"","Name":"","OntologyId":""},"TechnologyPlatform":"","CompatibleTools":[]},"DeviceEquipmentTechnologyData":{"CompatibleTools":[],"CellsTestedWith":[],"Placeholder":"Test"},"ServiceTechnologyData":{"CellsTestedWith":[],"Placeholder":"test"},"CodePipelineTechnologyData":{"Placeholder":"Test"}}}