Analysis of expression of immune system genes that are responsible for activation and inhibition of T-cell immune response in renal transplant recipients after extracorporeal photochemotherapy

• Ivan B. Kozlov
• Vatazin A.V.
• Kildushevsky A.V.
• Zulkarnaev A.B.
• Fedulkina V.A.
• Faenko A.P.
• Yazdovskiy V.V.
• Georgii O. Gudima
• Ilya A. Kofiadi

Abstract

Introduction. Due to the uprising need for kidney transplantation, there is a demand for new methods of preventing rejection of the transplant, based on the mechanisms of natural regulation of the immune response. One of the promising technologies in this area is extracorporeal photopheresis (ECP), based on electromagnetic influence on the recipient’s cells. The clinical efficacy of ECP has been confirmed by many studies, but the mechanism of therapeutic effect has not been sufficiently studied. Characterization of immune response during ECP is an important step towards formation of practical recommendations on clinical application of ECP and modification of immunosuppressive therapy.

The aim of this study was to analyze the changes in the expression level of the genes responsible for the activation and inhibition of the T-cell response in the recipients of the renal graft after ECP.

Material and methods. An open cohort randomized trial was conducted with 20 patients who underwent a single group cadaver kidney transplantation from an unrelated donor. The recipients of the compared groups received standard immunosuppressive therapy and the same therapy in combination with ECP procedures, according to the study protocol. The level of gene expression was analyzed by real-time RT-PCR. Comparison of the study groups was carried out in point 1 (1^st-4^th day) and point 2 (30^th day).

Results. The study of organ function in recipients of paired kidney allograft who received standard immunosupressive therapy and therapy in combination with ECP was carried out. The analysis of clinical data did not show statistically significant clinical differences, but in the specific consideration of each pair of cases the tendency to some improvement of the transplant organ engraftment rate in the early terms after the transplant, better survival of the graft and the recipient was revealed. The level of expression of genes involved in immune response regulation (CD28, CTLA4, PDL1, FOXP3) and cytokine genes (TNFA, IL1, IL2, IL10, IFNG) in the studied groups was determined.

Conclusion. According to the results of comparative analysis the level of expression of pro-inflammatory cytokines' genes IL1, IL2 and IFNG decreased to 30^th day after transplantation. A 30-day comparison between groups shows an increase in the expression level of the PDL1and FOXP3 genes (p = 0.0009 and 0.0013, correspondingly). Increased expression of these genes may be associated with the activation of regulatory T-cells and the development of peripheral tolerance.

Keywords:extracorporeal photopheresis; immune tolerance; expression level analysis; immune response regulation; regulatory T-cells

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