Evaluation of lymphocytes inactivation by extracorporeal photopheresis using tetrazolium salt based-assay

Transfus Apher Sci. 2015 Oct;53(2):242-5. doi: 10.1016/j.transci.2015.05.004. Epub 2015 May 27.

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.

Keywords: CFSE; ECP; Extracorporeal photopheresis; Flow cytometry; Tetrazolium salt.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • Apoptosis*
  • Cell Proliferation*
  • Female
  • Graft vs Host Disease / blood*
  • Graft vs Host Disease / therapy*
  • Humans
  • Lymphocyte Activation*
  • Lymphoma, T-Cell / blood
  • Lymphoma, T-Cell / therapy
  • Male
  • Photopheresis / methods*
  • Tetrazolium Salts / pharmacokinetics*

Substances

  • Tetrazolium Salts