Abstract only
e15220
Background: The purpose of the study was to evaluate the effect of IL-2 and INF-γ on the proliferation and phenotype of lymphocytes in patients with breast cancer (BC) after ...T-regs removal from the pool in vitro. Methods: Lymphocytes were isolated from the blood of 11 patients with stage II BC, T-regs were removed by immunomagnetic separation, and samples were cultured for 6 days with cytokines: IL-2 - 0.1/1 μg, INF-γ - 10 IU and their combinations. Results: The level of NK cells in the samples with IL-2 was maintained throughout the experiment, with IFN-γ - gradually decreased, and in the control (samples without stimulation) it decreased sharply. In experimental samples (IL-2, IL-2+IFN-γ), the amount of NKT remained at the initial level by day 6, and in the control it decreased by 2 times (p < 0.05). The number of CD335+ NK cells with high cytotoxicity against target cells increased after 3 days in the test samples with IL-2 by 5.2 times, and after 6 days - by 11.6 times in comparison with the control (p < 0.05). On day 6, the level of double-positive T cells increased by 1.8 times in co-culturing with IL-2+INF-γ (1 μg/10 IU), (p < 0.05). The percentage of double-negative T cells increased in the dynamics of cultivation from days 1 to 6, in some cases statistically significantly (IL-2 - 1 μg: by 2 times; IL-2+INF-γ - 0.1 μg/10 IU: by 1.8 times; p < 0.05). The percentage of CD4+CD38+ cells on day 6 of incubation with IL-2 at both doses increased by 1.8-2 times compared with the control, and CD8+CD38+ by 4-6 times. The percentage of CD4+HLA-DR+ cells after 6 days of cultivation with IL-2 increased by 6 times, and CD8+HLA-DR+ by 8-9 times. Cultivation with INF-γ reduced the stimulating effect, and in some cases it did not appear. The combined effect of IL-2 and INF-γ on day 6 led to an increase in the proportion of CD4+CD25+CD127dim cells by 1.8 times in comparison with the control (p < 0.05), and when exposed to INF-γ only, the number of these cells remained at the control levels. Conclusions: INF-γ is not an effective inducer of activation of immune effector cells, while the effect of IL-2, both alone and in combination with INF-γ, leads to a significant increase in the proportion of cytotoxic cells, as well as to activation of the T cell unit, including CD4 + CD25 + CD127dim.
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e15201
Background: A review of scientific literature has shown that IL-2 is most often used for the LAK generation, while the potential of other NK-stimulating interleukin cells remains ...poorly understood. In this study, we investigated the effect of IL-7 and IL-15 on ex vivo LAC generation. Methods: A fraction enriched in NK cells was isolated by magnetic cell sorting with the NK Cell Isolation Kit (#130-092-657, Miltenyi Biotec, Germany) from PBMC in 11 patients with stage II-III breast cancer without treatment. Cells were introduced into a 6-well 3x10
5
plate in RPMI medium (Gibco, USA) supplemented with 10% FBS (Gibco, USA). Cytokines 40 ng/ml were added to the wells in 6 variants: 1) IL-15; 2) IL-2; 3) IL-7; 4) IL-15+IL-7; 5) IL-15+IL-7+IL-2; 6) control without cytokines. Cells were cultured at 5.0% CO
2
and 37°C. Cells were counted with a hemocytometer daily for 5 days and on days 8, 9 and 10 of cultivation. Results: The number of NK cells in control samples gradually decreased: by 2 times on day 5 and by 3 times on day 10. On day 5, the number of NK cells was 1.5 times higher than in the control when cultured with IL-2, and 1.4 times higher when cultured with IL-7+IL-15. After 9 days, a statistically significant increase in the number of cells, compared to the control sample, was observed with the addition of IL-2 (1.6 times); IL-15 and IL-7+IL-15 (1.5 times). On day 10, significant differences from the control were found in most samples: the number of cells was higher in samples cultured with IL-2 and IL-7+IL-15 (1.9 times) and with IL-15 and IL-2+IL-7+IL-15 (1.7 times). IL-7 alone led to a gradual decrease in the number of cells, and on days 8, 9 and 10 it was lower than in the control samples. Conclusions: In general, the introduction of cytokines into the samples enriched with NK cells contributed to the preservation of this subpopulation on days 5-10 of cultivation. However, the use of IL-7 and IL-15, both alone and in combination, did not lead to a significant increase in LAK compared to the use of IL-2.
Aim
To study the practice of drug treatment of ischemic heart disease (IHD) and the consistency of this practice with the established guidelines.
Material and methods
Results of the Russian ...part of the EUROASPIRE V study were compared with the general European population of the study. At ≥6 mos. and <2 years after the discharge from the hospital, patients were invited to visit the site for an interview. The drug therapy recommended upon discharge and taken by patients in the long-term as well as the patients’ compliance with the treatment were analyzed. In Russian centers, 699 patients were registered, and 399 of them visited the centers for the interview.
Results
Upon discharge from the hospital, patients of the Russian cohort and of the entire study population were prescribed acetylsalicylic acid or other antiplatelet drugs (99.2% and 94.1%, respectively); beta-blockers (87.2 and 81.6%, respectively); angiotensin-converting enzyme (ACE) inhibitors (69.9% and 61.1%, respectively); sartans (16.5% and 14.2 %, respectively); calcium channel blockers (19.3 and 19.4 %, respectively); nitrates (8.0% and 22.5 %, respectively); diuretics (31.1 and 32.5 %, respectively); statins (98.0% and 85.0 %, respectively); and anticoagulants (6.6 and 8.3 %, respectively). For the long-term treatment, patients of the Russian cohort and of the entire study population took antiplatelets (94.7 % and 92.5 %, respectively); beta-blockers (83.2% and 81.0 %, respectively); ACE inhibitors (60.2% and 57.3 %, respectively); sartans (19.3% and 18.4 %, respectively); calcium antagonists (21.1% and 23.0 %, respectively); nitrates (9.0% and 18.2 %, respectively); diuretics (31.8% and 33.3 %, respectively); statins (88.2% and 80.8 %, respectively); and anticoagulants (8.8% and 8.2 %, respectively). High intensity hypolipidemic therapy was prescribed to 54.0 % of patients in Russian centers and 60.3 % of patients in the entire study. Both Russian and international patients evaluated their compliance with the prescribed medication as high.
Conclusion
According to results of the EUROASPIRE V study as compared to earlier studies, the practice of drug therapy in Russian patients with IHD has significantly approached European indexes. Further optimization is possible by a more extensive use of high intense hypolipidemic treatment and antidiabetic drugs with a documented positive effect on prognosis of cardiovascular diseases.
In this work we studied the effect of different concentrations of interleukin-2 (IL-2) and interferon-γ (IFN-γ) on the proliferation and immunophenotype of lymphocytes obtained from patients with ...locally advanced breast cancer (stage II-III) after immunomagnetic depletion of regulatory T cells (Tregs) from the general pool of lymphocytes in vitro. The peripheral blood of 11 patients was used as a material. Peripheral blood mononuclear cells (MNCs) were isolated and regulatory T lymphocytes were removed by immunomagnetic separation. After separation, the cells were cultured in a nutrient medium RPMI-1640 with 10% fetal bovine serum (FBS) for 7 days. Lymphocytes were activated on the first day of cultivation with one of the following cytokines: IFN-γ (10 IU/mL), IL-2 (0.1 or 1 μg/mL); IL-2 (0.1 or 1 μg/mL) and IFN-γ together. Lymphocytes without cytokine addition was used as a control. Cells were counted using an automatic counter before addition cytokines and after 2, 4, and 7 days of cultivation with cytokines, and their phenotype was examined. The results showed some phenotypic differences in some elements of cellular immunity between control and experimental samples. Particular attention is paid to the description of changes in the expression of surface markers in the natural killer (NK) subpopulation. It was noted that the proportion of Tregs, despite their preliminary depletion, increased after exposure to cytokines. As a result, the preliminary decrease in the proportion of Treg cells before the stimulation of lymphocytes did not produce the expected effect, therefore the use of depletion in such a methodological mode did not lead to significant results. However, one should not exclude the possibility of inhibiting Tregs by other methods.
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e13060
Background: Monotherapy with low doses of cytokines does not provide significant therapeutic results, while treatment with high doses leads to a number of side effects, for ...example, cytokine release syndrome, etc. Therefore, it is necessary to study the effect of cytokines on immune cells which are involved in the regulation of pro- and antitumor immune response. Thus, the aim of this study was to analyze the dynamics in the proliferation of the total fraction of lymphocytes after expansion and exposure to γc-cytokines: IL-2, IL-7 and IL-15 and their combinations in vitro in patients with diagnosed breast cancer. Methods: The study included 10 patients with locally advanced stage I-III breast cancer. Peripheral blood mononuclear cells (PBMC, ficoll-hypaque) were used as material. After density separation the cells were cultured at a dose of 500 thousand cells / ml in 6-well plates (Biofil, China) in RPMI 1640 (Gibco, USA) supplemented with 10% FBS (HyClone, USA) at 37° C, 5% CO2. Expansion of lymphocytes was performed by kit with anti-biotin magnetic particles MACSiBead to CD2, CD3 and CD28 (Miltenyi Biotec, Germany), according to the manufacturer's protocol. Stimulation of lymphocyte proliferation was performed on days 4, 8 and 12 by introducing recombinant cytokines - IL-2, IL-7, IL-15 (Miltenyi Biotec, Germany) and their combinations. Cytokines were introduced at a concentration of 40 ng / ml each in the following variants: IL-2; IL-2 / IL-15; IL-2 / IL-15 / IL-7; IL-15; IL-15 / IL-7. Plates with cell suspension were cultured at 37 ° C, 5% CO2 for 15 days. Results: The data obtained on the 11th day of incubation demonstrated statistically significant differences in cell viability in samples with the addition of interleukin IL-7 / IL-15 combination (778%) compared to control samples by 1.5 times (p < 0.05). The proportion of lymphocytes in samples with the addition of only IL-15 (702%) and IL-7 / IL-15 (756%) combination was 1.47 and 1.6 times higher, respectively, compared with the control (p < 0.05) at the 12th day of co-cultivation. Conclusions: Stimulation of lymphocyte proliferation by the IL-7 / IL-15 combination results in a high frequency of viable cell production. Despite the only culture stage, the results of the study may be important for optimizing the use of γc-cytokines in the treatment of patients with breast cancer.
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