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Biological quality control for extracorporeal photochemotherapy: Assessing mononuclear cell apoptosis levels in ECP bags of chronic GvHD patients
Author(s) -
Taverna Francesca,
Coluccia Paola,
Arienti Flavio,
Birolini Annalisa,
Terranova Laura,
Mazzocchi Arabella,
Rini Francesca,
Mariani Luigi,
Melani Cecilia,
Ravagnani Fernando
Publication year - 2015
Publication title -
journal of clinical apheresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.697
H-Index - 46
eISSN - 1098-1101
pISSN - 0733-2459
DOI - 10.1002/jca.21357
Subject(s) - medicine , apoptosis , extracorporeal photopheresis , propidium iodide , peripheral blood mononuclear cell , annexin , flow cytometry , immunology , graft versus host disease , programmed cell death , surgery , in vitro , transplantation , biology , biochemistry
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 (Δ APOPTOSIS ) after 24 h revealed the efficacy of the treatment. In 88.6% of the 31 ECP procedures, Δ APOPTOSIS was >15% (the “alerting” threshold for Δ APOPTOSIS 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. J. Clin. Apheresis 30:162–170, 2015. © 2014 Wiley Periodicals, Inc.