Since dendritic cells (DC) are involved in the development of autoimmune inflammation, researcher... more Since dendritic cells (DC) are involved in the development of autoimmune inflammation, researchers consider DC both as target cells for specific therapy of rheumatoid arthritis (RA) and as candidate cells for the development of cell-based methods to treat autoimmune diseases. The development of treatment strategies requires comprehensive research into the quantitative and qualitative characteristics of DC subtypes both ex vivo from RA patients and in vitro, to determine the possibility of inducing functionally mature DC in RA. To study the phenotypic and functional properties of myeloid (mDC) and plasmacytoid (pDC) DC isolated from the peripheral blood of patients with RA and induced in vitro. Blood samples were obtained from RA patients and healthy donors. Immature DC in the whole blood and in vitro induced DC were characterized by the positive expression of CD80, CD83, CCR7, IL-10, IL-4, IL-12 and IFN-α. R848 and lipopolysaccharide were used to determine DC maturation ability. From PBMCs of RA patients and health donors DCs with myeloid (imDC) and plasmacytoid (ipDC) phenotype were induced. The relative count of mDC in the peripheral blood between studied groups did not differ. pDC count was significantly lower for RA patients. DC from RA patients were characterized by low expression levels of CD80 and CD83 on both populations cells and high expression of CCR7 only on pDC. An increase in pDC producing IL-12 and IFN-α and a decrease in mDC and pDC producing IL-4 and IL-10 were shown in RA. imDC and ipDC obtained from RA patients according to their phenotype and cytokine profile did not differ from those obtained from healthy donors. There is an imbalance between subpopulations of DC in the peripheral blood of RA patients. DC of RA patients are less mature. The data suggest the involvement of DC in RA pathogenesis and confirm DC participation in balance shift towards Th1-type immune responses. At the same time, in vitro induced RA DC are phenotypically and functionally competent.
Advances in oncoimmunology related to the definition of the basic mechanisms of the formation of ... more Advances in oncoimmunology related to the definition of the basic mechanisms of the formation of antitumor immune response, as well as the opening of tumor-associated antigens recognized by immune cells, allowed to start developing ways to influence the effector cells of the immune system to generate effective antitumor cytotoxic response. We investigated the possibility to stimulate an antitumor response in a culture of mononuclear cells of breast cancer patients by dendritic cells transfected with HLA-A*02:01-restricted DNA constructs. We isolated dendritic cells from peripheral blood monocytes and delivered our constructs to these cells by magnetic transfection. Additionally, a series of experiments with loading of dendritic cells with autologous tumor cell lysate antigens was conducted. We have shown that dendritic cells transfected with the HLA-A*02:01-restricted DNA constructs are effective in inducing an antitumor response in a culture of mononuclear cells of breast cancer pa...
Bulletin of Experimental Biology and Medicine, 2007
Lysosomotropic composition of dialdehyde dextran and amphotericin B had a greater therapeutic eff... more Lysosomotropic composition of dialdehyde dextran and amphotericin B had a greater therapeutic effect in mice with systemic candidiasis compared to free amphotericin B. This composition normalized glucocorticoid function of the adrenal glands and decreased the severity of liver destruction at late terms of granulomatous inflammation.
Dendritic cells (DCs) loaded with tumor-associated antigens (TAA) are known to be the important a... more Dendritic cells (DCs) loaded with tumor-associated antigens (TAA) are known to be the important agents in antitumor response realization and still figure in lots of treatment schemes in cancer immunotherapy research. Here, we evaluated a cell-based protocol involving the use of original DNA constructs encoding the wide range of TAA epitopes expressed on different epithelial cancers. The constructs were transfected into ex-vivo-generated DCs of cancer patients (breast cancer, colorectal cancer, and non-small cell lung cancer). Direct cytotoxicity assay of effector cells, activated with the transfected DCs, showed a significant increase in the cytotoxicity against autologous tumor cells. The use of DNA-constructs encoding a large number of TAA’s for the in vitro DC loading to activate the T cell response could be a reliable and unified approach for immunotherapy and relapse prevention in patients with epithelial cancers.
For modulation of antitumor cytotoxic activity of mononuclear cells in vitro, autologous dendriti... more For modulation of antitumor cytotoxic activity of mononuclear cells in vitro, autologous dendritic cells loaded with tumor lysate antigens were cultured with peripheral blood mono-nuclear cells from patients with epithelial ovarian cancer in the presence or absence of IL-12 and IL-18. The effi ciency of modulation was evaluated by cytotoxic activity of mononuclear cells against autologous tumor cells, by the production of IFN-γ, IL-4, and by the count of perforin-containing lymphocytes. It was demonstrated that dendritic cells stimulated cytotoxic immune response in mononuclear cell culture. Maximum induction of cytotoxic activity of mononuclear cells was attained in case of dendritic cells combination with IL-12 and IL-18 (increased death of autologous tumor cells, accumulation of perforin-positive lymphocytes, enhanced production of IFN-γ).
Effect of antifungal preparation amphotericin B and its lysosomotropic composition with dialdehyd... more Effect of antifungal preparation amphotericin B and its lysosomotropic composition with dialdehyde-dextran on functional state of phagocytizing cells in the dynamics of granulo-matous inflammation induced by C. albicans was studied on CBA mice. A stimulating effect of amphotericin B on the production of reactive oxygen species by peritoneal and bone marrow phagocytes was observed, while lysosomotropic form of the antibiotic did not stimulate generation of oxygen radicals.
Significant effort has been devoted to developing effective cancer vaccines based on dendritic ce... more Significant effort has been devoted to developing effective cancer vaccines based on dendritic cells (DCs) loaded with various tumour antigens, including DNA constructs that carry sequences of tumour-associated antigens (TAAs). Such vaccines efficiently and selectively activate the T cell immune response. In this study, we describe a method to induce an antitumour immune response in mononuclear cell (MNC) cultures from colorectal cancer patients using DNA-transfected DCs encoding TAA epitopes of carcinoembryonic antigen, epithelial cell adhesion molecule and mucin 4. DCs were obtained from peripheral blood monocytes of colorectal cancer patients. Magnetic-assisted transfection was used to deliver the genetic constructs to DCs. To assess the potency of the immune response, the antitumour cytotoxic response was assessed by lymphocyte intracellular perforin and the MNC cytotoxic activity against autologous tumour cells. We showed that polyepitope DNA-transfected DCs enhanced MNC antitumour activity, increasing tumour cell death and the percentage of perforin-positive lymphocytes. In addition, DNA-transfected DCs elicited a cytotoxic response that was as efficient as that of tumour lysate-loaded DCs. Taken together, the data suggest that it is feasible to induce an antitumour immune response in colorectal MNCs using transfected DCs. Thus, the DNA construct reported in this study may potentially be used in therapeutic and prophylactic DC-based vaccines.
Lysosomotropic composition of dialdehyde dextran and amphotericin B had a greater therapeutic eff... more Lysosomotropic composition of dialdehyde dextran and amphotericin B had a greater therapeutic effect in mice with systemic candidiasis compared to free amphotericin B. This composition normalized glucocorticoid function of the adrenal glands and decreased the severity of liver destruction at late terms of granulomatous inflammation.
In CBA mice infected with C. albicans, phasic pattern of granulomatosis development was observed.... more In CBA mice infected with C. albicans, phasic pattern of granulomatosis development was observed. In all groups, the number of granulomas in the liver was minimum on day 56 after infection. Treatment with free amphotericin B and its composition with dialdehyde dextran (CA) reduced the number of infiltrations and granulomas in the liver, the changes were more pronounced in animals receiving CA. A different pattern of cyclic fluctuations of cortisol content in the blood and adrenal glands and progesterone content in the adrenal gland was observed. By the end of observation (day 84), cortisol content in the blood and adrenals of mice treated with CA was considerably lower than in untreated mice and animals receiving amphotericin B.
We compared the effects of dextrans with a molecular weight of 35 kDa oxidized by chemical (OD ch... more We compared the effects of dextrans with a molecular weight of 35 kDa oxidized by chemical (OD ch) and radiochemical (OD r) methods on oxidative and metabolic functions of peritoneal macrophages from BALB/c mice in vitro and in vivo. It was found that none type of dextrans exhibits chemiluminescent properties. In vitro study showed that OD ch had a priming effect on mouse peritoneal macrophages, while OD r did not poten-tiate the oxidative and metabolic response of cells to zymosan. Being injected intra-peritoneally, OD r more markedly enhanced chemiluminescent response of mouse peri-toneal macrophages and reduced their viability than OD ch. Thus, both dextrans are biocompatible, but in OD ch this parameter is higher.
Since dendritic cells (DC) are involved in the development of autoimmune inflammation, researcher... more Since dendritic cells (DC) are involved in the development of autoimmune inflammation, researchers consider DC both as target cells for specific therapy of rheumatoid arthritis (RA) and as candidate cells for the development of cell-based methods to treat autoimmune diseases. The development of treatment strategies requires comprehensive research into the quantitative and qualitative characteristics of DC subtypes both ex vivo from RA patients and in vitro, to determine the possibility of inducing functionally mature DC in RA. To study the phenotypic and functional properties of myeloid (mDC) and plasmacytoid (pDC) DC isolated from the peripheral blood of patients with RA and induced in vitro. Blood samples were obtained from RA patients and healthy donors. Immature DC in the whole blood and in vitro induced DC were characterized by the positive expression of CD80, CD83, CCR7, IL-10, IL-4, IL-12 and IFN-α. R848 and lipopolysaccharide were used to determine DC maturation ability. From PBMCs of RA patients and health donors DCs with myeloid (imDC) and plasmacytoid (ipDC) phenotype were induced. The relative count of mDC in the peripheral blood between studied groups did not differ. pDC count was significantly lower for RA patients. DC from RA patients were characterized by low expression levels of CD80 and CD83 on both populations cells and high expression of CCR7 only on pDC. An increase in pDC producing IL-12 and IFN-α and a decrease in mDC and pDC producing IL-4 and IL-10 were shown in RA. imDC and ipDC obtained from RA patients according to their phenotype and cytokine profile did not differ from those obtained from healthy donors. There is an imbalance between subpopulations of DC in the peripheral blood of RA patients. DC of RA patients are less mature. The data suggest the involvement of DC in RA pathogenesis and confirm DC participation in balance shift towards Th1-type immune responses. At the same time, in vitro induced RA DC are phenotypically and functionally competent.
Advances in oncoimmunology related to the definition of the basic mechanisms of the formation of ... more Advances in oncoimmunology related to the definition of the basic mechanisms of the formation of antitumor immune response, as well as the opening of tumor-associated antigens recognized by immune cells, allowed to start developing ways to influence the effector cells of the immune system to generate effective antitumor cytotoxic response. We investigated the possibility to stimulate an antitumor response in a culture of mononuclear cells of breast cancer patients by dendritic cells transfected with HLA-A*02:01-restricted DNA constructs. We isolated dendritic cells from peripheral blood monocytes and delivered our constructs to these cells by magnetic transfection. Additionally, a series of experiments with loading of dendritic cells with autologous tumor cell lysate antigens was conducted. We have shown that dendritic cells transfected with the HLA-A*02:01-restricted DNA constructs are effective in inducing an antitumor response in a culture of mononuclear cells of breast cancer pa...
Bulletin of Experimental Biology and Medicine, 2007
Lysosomotropic composition of dialdehyde dextran and amphotericin B had a greater therapeutic eff... more Lysosomotropic composition of dialdehyde dextran and amphotericin B had a greater therapeutic effect in mice with systemic candidiasis compared to free amphotericin B. This composition normalized glucocorticoid function of the adrenal glands and decreased the severity of liver destruction at late terms of granulomatous inflammation.
Dendritic cells (DCs) loaded with tumor-associated antigens (TAA) are known to be the important a... more Dendritic cells (DCs) loaded with tumor-associated antigens (TAA) are known to be the important agents in antitumor response realization and still figure in lots of treatment schemes in cancer immunotherapy research. Here, we evaluated a cell-based protocol involving the use of original DNA constructs encoding the wide range of TAA epitopes expressed on different epithelial cancers. The constructs were transfected into ex-vivo-generated DCs of cancer patients (breast cancer, colorectal cancer, and non-small cell lung cancer). Direct cytotoxicity assay of effector cells, activated with the transfected DCs, showed a significant increase in the cytotoxicity against autologous tumor cells. The use of DNA-constructs encoding a large number of TAA’s for the in vitro DC loading to activate the T cell response could be a reliable and unified approach for immunotherapy and relapse prevention in patients with epithelial cancers.
For modulation of antitumor cytotoxic activity of mononuclear cells in vitro, autologous dendriti... more For modulation of antitumor cytotoxic activity of mononuclear cells in vitro, autologous dendritic cells loaded with tumor lysate antigens were cultured with peripheral blood mono-nuclear cells from patients with epithelial ovarian cancer in the presence or absence of IL-12 and IL-18. The effi ciency of modulation was evaluated by cytotoxic activity of mononuclear cells against autologous tumor cells, by the production of IFN-γ, IL-4, and by the count of perforin-containing lymphocytes. It was demonstrated that dendritic cells stimulated cytotoxic immune response in mononuclear cell culture. Maximum induction of cytotoxic activity of mononuclear cells was attained in case of dendritic cells combination with IL-12 and IL-18 (increased death of autologous tumor cells, accumulation of perforin-positive lymphocytes, enhanced production of IFN-γ).
Effect of antifungal preparation amphotericin B and its lysosomotropic composition with dialdehyd... more Effect of antifungal preparation amphotericin B and its lysosomotropic composition with dialdehyde-dextran on functional state of phagocytizing cells in the dynamics of granulo-matous inflammation induced by C. albicans was studied on CBA mice. A stimulating effect of amphotericin B on the production of reactive oxygen species by peritoneal and bone marrow phagocytes was observed, while lysosomotropic form of the antibiotic did not stimulate generation of oxygen radicals.
Significant effort has been devoted to developing effective cancer vaccines based on dendritic ce... more Significant effort has been devoted to developing effective cancer vaccines based on dendritic cells (DCs) loaded with various tumour antigens, including DNA constructs that carry sequences of tumour-associated antigens (TAAs). Such vaccines efficiently and selectively activate the T cell immune response. In this study, we describe a method to induce an antitumour immune response in mononuclear cell (MNC) cultures from colorectal cancer patients using DNA-transfected DCs encoding TAA epitopes of carcinoembryonic antigen, epithelial cell adhesion molecule and mucin 4. DCs were obtained from peripheral blood monocytes of colorectal cancer patients. Magnetic-assisted transfection was used to deliver the genetic constructs to DCs. To assess the potency of the immune response, the antitumour cytotoxic response was assessed by lymphocyte intracellular perforin and the MNC cytotoxic activity against autologous tumour cells. We showed that polyepitope DNA-transfected DCs enhanced MNC antitumour activity, increasing tumour cell death and the percentage of perforin-positive lymphocytes. In addition, DNA-transfected DCs elicited a cytotoxic response that was as efficient as that of tumour lysate-loaded DCs. Taken together, the data suggest that it is feasible to induce an antitumour immune response in colorectal MNCs using transfected DCs. Thus, the DNA construct reported in this study may potentially be used in therapeutic and prophylactic DC-based vaccines.
Lysosomotropic composition of dialdehyde dextran and amphotericin B had a greater therapeutic eff... more Lysosomotropic composition of dialdehyde dextran and amphotericin B had a greater therapeutic effect in mice with systemic candidiasis compared to free amphotericin B. This composition normalized glucocorticoid function of the adrenal glands and decreased the severity of liver destruction at late terms of granulomatous inflammation.
In CBA mice infected with C. albicans, phasic pattern of granulomatosis development was observed.... more In CBA mice infected with C. albicans, phasic pattern of granulomatosis development was observed. In all groups, the number of granulomas in the liver was minimum on day 56 after infection. Treatment with free amphotericin B and its composition with dialdehyde dextran (CA) reduced the number of infiltrations and granulomas in the liver, the changes were more pronounced in animals receiving CA. A different pattern of cyclic fluctuations of cortisol content in the blood and adrenal glands and progesterone content in the adrenal gland was observed. By the end of observation (day 84), cortisol content in the blood and adrenals of mice treated with CA was considerably lower than in untreated mice and animals receiving amphotericin B.
We compared the effects of dextrans with a molecular weight of 35 kDa oxidized by chemical (OD ch... more We compared the effects of dextrans with a molecular weight of 35 kDa oxidized by chemical (OD ch) and radiochemical (OD r) methods on oxidative and metabolic functions of peritoneal macrophages from BALB/c mice in vitro and in vivo. It was found that none type of dextrans exhibits chemiluminescent properties. In vitro study showed that OD ch had a priming effect on mouse peritoneal macrophages, while OD r did not poten-tiate the oxidative and metabolic response of cells to zymosan. Being injected intra-peritoneally, OD r more markedly enhanced chemiluminescent response of mouse peri-toneal macrophages and reduced their viability than OD ch. Thus, both dextrans are biocompatible, but in OD ch this parameter is higher.
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Papers by Vasily Kurilin