title: Biological quality control for extracorporeal photochemotherapy: Assessing mononuclear cell apoptosis levels in ECP bags of chronic GvHD patients.
abstract: Extracorporeal photochemotherapy (ECP) is a treatment approved by the FDA for cutaneous T-cell lymphoma, and it is currently used off-label for graft-versus-host disease (GvHD) and other conditions. In agreement with good practices for the therapeutic use of human cells, quality control has to be performed to validate the ECP procedure with the off-line technique. Since no gold-standard biological test is available, we assessed the apoptosis generated in the ECP bag using a flow cytometric analysis. Thirty-one ECP procedures performed on 13 patients with chronic GvHD were studied by monitoring the induction of mononuclear cell (MNC) apoptosis using annexin V/propidium iodide double staining; residual lymphocyte proliferation to standard mitogens was also measured in 17 of the procedures. The kinetics of apoptosis was analyzed at different times in MNCs untreated or treated with 8-methoxy-psoralen plus ultraviolet A; the variation (ΔAPOPTOSIS ) after 24 h revealed the efficacy of the treatment. In 88.6% of the 31 ECP procedures, ΔAPOPTOSIS was >15% (the "alerting" threshold for ΔAPOPTOSIS was set at 15% on the basis of our data); in the remainder (19.4%), the increment in apoptosis was lower. In four procedures, the proliferation assay was useful for assessing the effect of ECP on the apheretic bag. In conclusion, both flow cytometric assays enabled a biologically significant result to be obtained. In our opinion, the apoptosis test-being faster and easier than the proliferation test-could be a reliable way to validate ECP procedures.
authors.affiliation: Immunohematology and Transfusion Medicine Service, Fondazione IRCCS, Istituto Nazionale dei Tumori, Milan, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Taverna
authors.affiliation: Immunohematology and Transfusion Medicine Service, Fondazione IRCCS, Istituto Nazionale dei Tumori, Milan, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Coluccia
authors.affiliation: Immunohematology and Transfusion Medicine Service, Fondazione IRCCS, Istituto Nazionale dei Tumori, Milan, Italy.
authors.forename: Flavio
authors.initials: F
authors.lastname: Arienti
authors.affiliation: Immunohematology and Transfusion Medicine Service, Fondazione IRCCS, Istituto Nazionale dei Tumori, Milan, Italy.
authors.forename: Annalisa
authors.initials: A
authors.lastname: Birolini
authors.affiliation: Immunohematology and Transfusion Medicine Service, Fondazione IRCCS, Istituto Nazionale dei Tumori, Milan, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Terranova
authors.affiliation: Immunohematology and Transfusion Medicine Service, Fondazione IRCCS, Istituto Nazionale dei Tumori, Milan, Italy.
authors.forename: Arabella
authors.initials: A
authors.lastname: Mazzocchi
authors.affiliation: Human Tumor Immunotherapy Unit, Fondazione IRCCS, Istituto Nazionale dei Tumori, Milan, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Rini
authors.affiliation: Clinical Epidemiology and Trial Organization Unit, Fondazione IRCCS, Istituto Nazionale dei Tumori, Milan, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Mariani
authors.affiliation: Scientific Directorate, Fondazione IRCCS, Istituto Nazionale dei Tumori, Milan, Italy.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Melani
authors.affiliation: Immunohematology and Transfusion Medicine Service, Fondazione IRCCS, Istituto Nazionale dei Tumori, Milan, Italy.
authors.forename: Fernando
authors.initials: F
authors.lastname: Ravagnani
mesh_terms: D000328:Adult; D000368:Aged; D017209:Apoptosis; D001781:Blood Component Removal; D049109:Cell Proliferation; D002469:Cell Separation; D005260:Female; D005434:Flow Cytometry; D006086:Graft vs Host Disease / Q000628:therapy*; D006801:Humans; D007700:Kinetics; D007937:Leukapheresis / Q000379:methods*; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000473:pathology; D008214:Lymphocytes / Q000166:cytology; D016410:Lymphoma, T-Cell, Cutaneous / Q000628:therapy; D008297:Male; D008730:Methoxsalen / Q000008:administration & dosage; D008875:Middle Aged; D017893:Photopheresis / Q000379:methods*; D011786:Quality Control; D015203:Reproducibility of Results; D019172:Transplantation Conditioning; D014466:Ultraviolet Rays
chemical_list: D008730:Methoxsalen
keywords: CFSE; ECP; annexin V/PI; flow cytometry