The effect of cryopreservation on the parameters of mononuclear apoptosis during extracorporeal photopheresis

Introduction. The mechanism of action of extracorporeal photopheresis (ECP) is associated with the induction of apoptosis of lymphocytes, which cause the activation of antigen-presenting cells (APC). As a result of the use of ECF, the number of T-regulatory lymphocytes increases, which induces an immunosuppressive effect. The actual problem lies in the use of cryopreserved and photo-irradiated mononuclear cells.

Aim — to analyze the functional features of cryopreserved mononuclear cells after ECP.

Materials and methods. The study analyzed the indicators of early and late stages of apoptosis in the concentrate of mononuclear cells of different groups, depending on the time of collection and the type of exposure on these concentrates of mononuclear cells: immediately after apheresis (groups 1.1 and 1.2), and after ECP (groups 1.3, 2.1 and 2.2), groups 1.2 and 2.2 were after cryopreservation, and 1.3 — after cryopreservation, and then after ECP. 113 samples of mononuclear cell concentrates obtained from 12 patients with chronic graft-versus-host reaction (GVHD) were analyzed. All groups of samples were cultured to determine the dynamics of changes in indicators of early and late stages of apoptosis over time.

Results. The proportion of lymphocytes in the late stage of apoptosis after 2 days cultivation, after the standard ECP procedure, was comparable to the proportion of lymphocytes in the late stage of apoptosis during ECP followed by cryopreservation of the leukoconcentrate.

Conclusion. It is advisable to collect mononuclear cells, perform their extracorporeal photo-irradiation, and then divide the photo-irradiated mononuclear cells into several parts both for cryopreservation and for the return of non-cryopreserved photo-irradiated mononuclear cells to the patient.

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