title: Flow cytometric evaluation of lymphocyte transformation test based on 5-ethynyl-2'deoxyuridine incorporation as a clinical alternative to tritiated thymidine uptake measurement.
abstract: In clinical laboratories, the evaluation of lymphocyte proliferative response (lymphocyte transformation test-LTT) is routinely performed by the measurement of [(3)H]-thymidine uptake after stimulation. In this study we evaluated the performances of a recently developed non-radioactive test based on the detection by flow cytometry of 5-ethynyl-2'deoxyuridine (EdU) incorporation for the measurement of LTT in routine lab conditions. After definition of optimal protocol parameters, EdU incorporation test showed good repeatability and reproducibility. Moreover, this assay was flexible enough to fit important clinical laboratory constraints (delayed stimulation, low number of cells and delayed analysis after staining). Importantly, correlations between results obtained with EdU and [(3)H]-thymidine incorporation assays were excellent both in healthy volunteers and pediatric and septic patients. In particular, the two techniques identified patients presenting with altered LTT. Upon confirmation in a larger cohort of patients, EdU incorporation assay may be a relevant non-radioactive candidate for LLT in clinic.
authors.affiliation: BioMérieux, Joint Research Unit, bioMérieux-Hospices Civils de Lyon, Hôpital Edouard Herriot, Lyon, France. Electronic address: fanny.poujol@biomerieux.com.
authors.forename: Fanny
authors.initials: F
authors.lastname: Poujol
authors.affiliation: BioMérieux, Joint Research Unit, bioMérieux-Hospices Civils de Lyon, Hôpital Edouard Herriot, Lyon, France; Cellular Immunology Laboratory, Hôpital Edouard Herriot, Hospices Civils de Lyon, Lyon, France. Electronic address: guillaume.monneret@chu-lyon.fr.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Monneret
authors.affiliation: Intensive Care Units, Centre Hospitalier Lyon-Sud, Hospices Civils de Lyon, Lyon, France. Electronic address: arnaud.friggeri@chu-lyon.fr.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Friggeri
authors.affiliation: Intensive Care Units, Hôpital Edouard Herriot, Hospices Civils de Lyon, Lyon, France. Electronic address: thomas.rimmele@chu-lyon.fr.
authors.forename: Thomas
authors.initials: T
authors.lastname: Rimmelé
authors.affiliation: Cellular Immunology Laboratory, Hôpital Edouard Herriot, Hospices Civils de Lyon, Lyon, France. Electronic address: christophe.malcus@chu-lyon.fr.
authors.forename: Christophe
authors.initials: C
authors.lastname: Malcus
authors.affiliation: Cellular Immunology Laboratory, Hôpital Edouard Herriot, Hospices Civils de Lyon, Lyon, France. Electronic address: francoise.poitevin@chu-lyon.fr.
authors.forename: Françoise
authors.initials: F
authors.lastname: Poitevin-Later
authors.affiliation: BioMérieux, Joint Research Unit, bioMérieux-Hospices Civils de Lyon, Hôpital Edouard Herriot, Lyon, France. Electronic address: alexandre.pachot@biomerieux.com.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Pachot
authors.affiliation: Intensive Care Units, Centre Hospitalier Lyon-Sud, Hospices Civils de Lyon, Lyon, France. Electronic address: alain.lepape@chu-lyon.fr.
authors.forename: Alain
authors.initials: A
authors.lastname: Lepape
authors.affiliation: BioMérieux, Joint Research Unit, bioMérieux-Hospices Civils de Lyon, Hôpital Edouard Herriot, Lyon, France; Intensive Care Units, Hôpital Edouard Herriot, Hospices Civils de Lyon, Lyon, France. Electronic address: julien.textoris@biomerieux.com.
authors.forename: Julien
authors.initials: J
authors.lastname: Textoris
authors.affiliation: BioMérieux, Joint Research Unit, bioMérieux-Hospices Civils de Lyon, Hôpital Edouard Herriot, Lyon, France; Cellular Immunology Laboratory, Hôpital Edouard Herriot, Hospices Civils de Lyon, Lyon, France. Electronic address: fabienne.venet@chu-lyon.fr.
authors.forename: Fabienne
authors.initials: F
authors.lastname: Venet
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D001692:Biological Transport; D016022:Case-Control Studies; D049109:Cell Proliferation / Q000187:drug effects; D003857:Deoxyuridine / Q000031:analogs & derivatives* / Q000276:immunology / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes; D006801:Humans; D008213:Lymphocyte Activation / Q000187:drug effects*; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D010798:Phycocyanin; D010835:Phytohemagglutinins / Q000494:pharmacology; D061251:Primary Cell Culture; D015203:Reproducibility of Results; D012772:Shock, Septic / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D013936:Thymidine / Q000276:immunology / Q000378:metabolism; D014316:Tritium
chemical_list: D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D010835:Phytohemagglutinins; C002497:allophycocyanin; D014316:Tritium; D010798:Phycocyanin; C031086:5-ethynyl-2'-deoxyuridine; D013936:Thymidine; D003857:Deoxyuridine
keywords: EdU; Flow cytometry; Lymphocyte; Proliferation; [(3)H]-Thymidine