{"doi":"10.3389/fimmu.2020.00633","title":"Multiplex T Cell Stimulation Assay Utilizing a T Cell Activation Reporter-Based Detection System","abstract":"Recent advancements in single cell sequencing technologies allow for identification of numerous immune-receptors expressed by T cells such as tumor-specific and autoimmune T cells. Determining antigen specificity of those cells holds immense therapeutic promise. Therefore, the purpose of this study was to develop a method that 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.","journal":"Frontiers in Immunology","year":2020,"id":92802,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":41,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.959,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":277806,"name":"Zhicheng Zhou","orcid":"0000-0001-9137-7951","position":1,"is_corresponding":false},{"id":463865,"name":"Laurie G. Landry","orcid":"0000-0002-4064-9109","position":2,"is_corresponding":false},{"id":463866,"name":"A. Anderson","orcid":"0000-0001-9746-1257","position":3,"is_corresponding":false},{"id":464704,"name":"Aimon K. Alkanani","orcid":null,"position":4,"is_corresponding":false},{"id":463867,"name":"J Fischer","orcid":"0000-0003-0001-4384","position":5,"is_corresponding":false},{"id":393227,"name":"Mark Peakman","orcid":"0000-0003-3328-3513","position":6,"is_corresponding":false},{"id":457125,"name":"Roberto Mallone","orcid":"0000-0002-9846-8861","position":7,"is_corresponding":false},{"id":452316,"name":"Kristen Campbell","orcid":null,"position":8,"is_corresponding":false},{"id":463868,"name":"Aaron W. Michels","orcid":"0000-0003-3766-5244","position":9,"is_corresponding":false},{"id":369808,"name":"Maki Nakayama","orcid":"0000-0002-0009-3687","position":10,"is_corresponding":false},{"id":464703,"name":"Sarah Mann","orcid":null,"position":0,"is_corresponding":true}],"reference_count":59,"raw_metadata":null,"created_at":"2026-07-18T22:30:31.563852Z","pmid":"32328071","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}