title: Evaluation of lymphocytes inactivation by extracorporeal photopheresis using tetrazolium salt based-assay.
abstract: Extracorporeal photopheresis (ECP) is accepted as a second-line therapy for the treatment of acute and chronic steroid-refractory graft versus host disease (GvHD), cutaneous T-cell lymphoma and solid organ transplantation. ECP should be validated: we compared in parallel apoptosis and proliferation analysis of patient lymphocytes treated with 8-MOP ECP using respectively Annexin V/7-aminoactinomycin D (7-AAD) and CFSE with a tetrazolium salt (WST-1) method. Using WST-1 assay we found a significant decrement (p < 0.01) of metabolic activity at 4 days between ECP-treated and untreated cells. This finding was confirmed by the significant decrease of cell proliferation and increase of cell death observed by CFSE and 7AAD-Annexin V, respectively. Accordingly, once validated against a reference method, WST-1 could represent a rapid and easy assay for routinely quality control of ECP.
authors.affiliation: Fondazione Progetto Ematologia (Hematology Project Foundation), Vicenza, Italy.
authors.forename: Katia
authors.initials: K
authors.lastname: Chieregato
authors.affiliation: Immunohematology and Transfusion Medicine Service, DIMT, San Bortolo Hospital, Vicenza, Italy.
authors.forename: Alberta
authors.initials: A
authors.lastname: Alghisi
authors.affiliation: Department of Cellular Therapy and Hematology, San Bortolo Hospital, Vicenza, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Borghero
authors.affiliation: Department of Cellular Therapy and Hematology, San Bortolo Hospital, Vicenza, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Elice
authors.affiliation: Department of Cellular Therapy and Hematology, San Bortolo Hospital, Vicenza, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Raimondi
authors.affiliation: Immunohematology and Transfusion Medicine Service, DIMT, San Bortolo Hospital, Vicenza, Italy.
authors.forename: Ermella
authors.initials: E
authors.lastname: Zanetti
authors.affiliation: Fondazione Progetto Ematologia (Hematology Project Foundation), Vicenza, Italy; Department of Cellular Therapy and Hematology, San Bortolo Hospital, Vicenza, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Rodeghiero
authors.affiliation: Department of Cellular Therapy and Hematology, San Bortolo Hospital, Vicenza, Italy. Electronic address: astori@hemato.ven.it.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Astori
mesh_terms: D017209:Apoptosis; D049109:Cell Proliferation; D005260:Female; D006086:Graft vs Host Disease / Q000097:blood* / Q000628:therapy*; D006801:Humans; D008213:Lymphocyte Activation; D016399:Lymphoma, T-Cell / Q000097:blood / Q000628:therapy; D008297:Male; D017893:Photopheresis / Q000379:methods*; D013778:Tetrazolium Salts / Q000493:pharmacokinetics*
chemical_list: D013778:Tetrazolium Salts
keywords: CFSE; ECP; Extracorporeal photopheresis; Flow cytometry; Tetrazolium salt