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Information × Registration Number 0217U004788, 0116U002409 , R & D reports Title To develop and justify the feasibility of modern nano- and cellular technologies to enhance the anticancer and immunomodulatory properties of vaccines based on dendritic cells popup.stage_title Head Khranovska Natalia, Registration Date 03-03-2017 Organization Ukrainian Research Institute of Oncology and Radiology popup.description2 Object of the study - phenotypic and functional characteristics of dendritic cells (DC) of patients with malignant tumors and healthy people, technologies of cancer vaccines based on DC design, cytokine mRNA expression level. Aim - to develop an effective technology for obtaining of Tx-1 - polarizing DC monocytic using different bioactive mediators. Methods - cell cultures, flow cytometry, immunological, molecular genetic, statistics. Equipment - FACSCalibur flow cytometer, centrifuge OPN-3, microscope, scales WPS 600/C and W, Thermocycler Veriti 96 Well Thermal Cycler, fluorescent microscope Imager.M1 "Axio Zeizz", Germany, spectrophotometer Nano Drop 1000, 7500 Real-Time PCR Systems. It was established that DCs generated from monocytes of patients with malignancies with their cytomorphological for phenotypic properties were related to medium maturity cell, express high levels of Bcl-2 mRNA and CCR7 comparing to DCs obtained from healthy people. It was established that the level of mRNA expression of immunosuppressive cytokine TGF-b, IL-10 and the enzyme IDO in generated DCs of patients with malignant tumors was increased comparing to healthy people DCs (p> 0.05) and the level of IL-12 bioactive production was also reduced. It was proved that the combination of two activating signals - IFN-a and LPS did not significantly affected the phenotypic characteristics compared with DC activation only via LPS. It was established that the combination of LPS + IFN-a promotes Th1 - polarization of generated DCs, while not inducing their tolerogeneic properties. Using IFN-a (10 thousand. IU / ml) an increase of TNF-a mRNA expression (p> 0.05) and a slight reduction of mRNA of IL-10 expression was observed; increase of chemokines RANTES and MIP 1-a 2-7 mRNA expression times (p> 0.05) was observed. It was found that cytoplasmic membrane extract (CPM) of Staphylococcus aureus Wood 46 in the lowest used concentrations (0.2 mg/ml) contributed to significant increase of CD86 and HLA-DR molecules expression in the generated DCs. Staphylococcus aureus Wood 46 CPM significantly affected the phagocytic activity of the generated DCs, and thus had a dose-dependent effect: concentration of CPM increasing decreased the phagocytic activity of DCs. It was proved that the combination of IFN-a (10 thousand IU/ml) and Staphylococcus aureus Wood 46 CPM promoted mRNA IFN-a and TNF-a Th1-polarizing cytokines expression in the generated DCs (p> 0.05), and resulted in significant increase of the level of CCR7 chemokine expression, (p> 0.05), indicating that the increase in DC migration activity and the ability to initiate immune response. Our results showed the possibility and potential of the combination of factors IFN-a and LPS and IFN-a and Staphylococcus aureus Wood 46 CPM for methods optimization to obtain mature DCs, suitable for use as cellular basis for the creation of individual therapeutic cancer vaccines. Product Description popup.authors Іномістова М.В. Сидор Р.І. Скачкова О.В. Храновська Н.М. popup.nrat_date 2020-04-02 Close