APPLICATION OF LEUKOCYTE FILTERS TO OBTAIN A PREPARATION OF CYTOKINE-INDUCED KILLER CELLS

Abstract

The ability of CIK cells to recognize and lyse tumor cells has been demonstrated in preclinical studies in vitro. The ClinicalTrials.gov portal has registered over 600 completed clinical trials devoted to the method of cellular immunotherapy, including over 30 on the use of CIK. There are many ways to obtain cells drugs based on lymphocytes. Their common drawback is the need for additional blood sampling to obtain a cell preparation in large volumes, so it remains relevant to develop new approaches that take into account the solution to this problem. The purpose of the study was to evaluate the possibility of using leukocyte filters as a source of lymphocytes to obtain large doses of CIK. Materials and methods. The article describes in detail the procedure for extracting lymphocytes from leukocyte filters (30 donors). Lymphocytes were cultured in a CO2 incubator for 10-14 days in a medium with IL-2 and IL-15. The level of cell activation was assessed by ELISA and flow cytometry. The subpopulation composition of T, NK, TNK, B lymphocytes (CD3, CD4, CD8, CD14, CD16, CD19, CD45, CD56) and activation markers on them (CD38 and HLA-DR) were assessed before and after activation. Results. The possibility of using leukocyte filters after separation of plasma with leukocytes remaining on them was assessed. The functional activity of extracted lymphocytes and the cytokine composition of supernatants after cultivation in a complete nutrient medium were studied. Peptide complex (supernatant) contained high doses of signaling proteins: IL-2, IL-6 IL-10, IFNγ, TNFα and retained its functional activity after long-term storage at low temperatures. The drug is sterile and consists predominantly of T-, TNK- and NK- cells with a high level of viability and contains activation markers: HLA-DR, CD38. Conclusion.  Thus, up to 350 million activated lymphocytes and aliquots with the supernatant were obtained from one filter. The drug can be used both for basic research and for administering AIT to cancer patients in order to restore the patients’ own antitumor immunity potential. This amount of cellular and acellular preparation is sufficient to conduct a larger number of AIT courses without additional blood sampling.

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