Introduction. The increasing clinical use of cellular technologies suggests the possibility of long-term storage of the cellular product while maintaining its viability and basic properties. The procedures of cell’s cryopreservation used in laboratory as well in clinical practice differ a lot. Each method includes two tasks to solve: what is the optimal freezing medium to use and what cryopreservation procedure to prefer. In this paper, we present the method utilized in our center for bone marrow cell cryopreservation. The freezing was carried out in nitrogen vapor after adding the medium containing 95 % dextran [average mw 50 000–70 000] and 5 % dimethylsulfoxid.Purpose. To show that the proposed method of cryopreservation of dendritic cells is highly effective, simple, reproducible and most convenient for clinical use.Materials and methods. Viability, expression of surface antigens and stimulating activity towards allogeneic T lymphocytes of cryopreserved mature dendritic cells cultured from peripheral blood monocytes were evaluated.Results. The first cryopreservation resulted in the death of a small amount of cells. The second freezing procedure increased the proportion of dead cells. Meanwhile, the difference in the expression of the surface antigens in fresh, cryopreserved and re-cryopreserved dendritic cells was not statistically significant. The level of stimulating activity of fresh and cryopreserved dendritic cells did not significantly differ. Conversely the proliferation of allogeneic T lymphocytes was decreased after stimulation with re-cryopreserved dendritic cells.Conclusion. The presented method of cryopreservation allows to preserve the viability and basic functions of dendritic cells. After thawing dendritic vaccine could be administered to patients after being diluted in an isotonic saline without washing, which makes this method the most convenient for clinical use.
Background. The incidence of oral mucosa cancer (OMC) is higher in people over 50 years of age, and the aggressiveness of the course of the disease is higher in people under 50 years of age. In this context, it is of interest to clarify the mechanisms of immune disorders characteristic of patients of different age groups.Aim. To research systemic and local immunity in OMC patients and the relationship of peripheral blood lymphocyte population (PBLs) and tumor infiltrating lymphocytes (TILs) with the patient’s sex and age.Materials and methods. PBLs and TILs effector and suppressor populations were studied by flow cytometry in OMC patients aged 29 to 84 years.Results. The percentage of CD3-, CD3+CD4+ and CD3+CD8+T cells, regulatory CD4+CD25+CD127low/ –(CD4Treg) and CD8+CD11b–CD28–(CD8Тreg) T lymphocytes, CD4+PD-1+ and CD8+PD-1+ T cells was increased in TILs compared to PBLs. The levels of cytotoxic CD8+CD11b+CD28– T lymphocytes, NK, CD8+Perforin+ and CD16+Perforin+ cells in TILs were lower than in PBLs. The relationship between the level of CD4Treg and other TILs and PBLs depended on the patient’s sex. Age-related changes in the levels of NK and CD8 T-cells were observed in men, and CD4Treg – in women.Conclusion. Local immunity in OMC patients is highly immunosuppressive. The sex of patients influences the relationship between CD4Treg and other populations of PBLs and TILs, as well as age-related changes in the OMC patients’ immune system. This investigation results can make a certain contribution to personalized treatment of patients with OMC, taking into account differences in systemic and local immunity and in the immune response to the tumor in patients of different sex and age.
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