Considering that both innate and adaptive immune responses are involved in the pathogenesis of Crohn's disease(CD), novel therapeutic options have significantly been developed. Biological agents represent an impor...Considering that both innate and adaptive immune responses are involved in the pathogenesis of Crohn's disease(CD), novel therapeutic options have significantly been developed. Biological agents represent an important addition to the conventional treatments for immuno-inflammatory conditions, acting as antagonists of adhesion molecules or various inflammatory cytokines. The interleukin 12(IL-12)/IL-23 common pathway has been found to play a determinant role in the induction of inflammation in adaptive immune responses. In particular, IL-23 promotes the differentiation of na?ve T helper cells into Th17 phenotype with the concomitant secretion of several inflammatory cytokines such as IL-17 and IL-22, whereas IL-12 induces the Th1 polarization and production of critical cytokines such as interferon-γ and tumor necrosis factor. Nowadays, there is increased interest regarding the role of IL-23 as a therapeutic target of CD through the blockage of IL-23 mediated pathways. In this editorial, we focus on the role of IL-12/IL-23 pathway in the regulation of mucosal immunity and in the induction and maintenance of chronic inflammation. In parallel, we critically discuss the available data regarding the therapeutic effect of the IL-12/IL-23 inhibitors and especially of ustekinumab, a human monoclonal antibody which has been recently approved by the United States Food and Drug Administration for the management of moderateto-severe CD and its potential to be used as first-line therapy in everyday clinical practice.展开更多
The full-length cDNA encoding the subunits p40 and p35 of human interleukin12(hIL12) were cloned separately by RTPCR, linked together by internal ribosomal entry site (IRES) of encephalomyocarditis virus which initiat...The full-length cDNA encoding the subunits p40 and p35 of human interleukin12(hIL12) were cloned separately by RTPCR, linked together by internal ribosomal entry site (IRES) of encephalomyocarditis virus which initiates capindependent translation to form a dicistronic gene fragment. The dicistronic fragment was placed between the cytomegalovirus (CMV) promoter and SV40 polyA signal to form a dicistronic expression cassette. Subsequently, the dicistronic expression cassette was inserted into E1 region of Ad5 genome in cosmid vector pAx1cw of E1substitution type. By homologous recombination with EcoT22Idigested Ad5 DNATPC in 293 cells, the replicationdeficient recombinant adenoviruses of hIL12 were generated efficiently. After infected with hIL12 recombinant adenoviruses in vitro, 293 cells, human hepatocellular carcinoma cells HepG2, and primary human skin fibroblasts expressed and secreted hIL12 at comparable levels (30～60ng/ 106cells/24hr), which could stimulate the proliferation and IFNγ production of human lymphoblasts. These suggest that the dicistronic adenovirus vector of hIL12 could effectively mediate the expression of bioactive hIL12 and might be used in cancer gene therapy.展开更多
丙型肝炎病毒（hepatitis C virus,HCV）是导致慢性肝炎的主要病原之一^（[1]）,HCV感染后20%～30%患者能自主清除,近70%可发展为慢性感染^（[2]）。免疫应答过程中分泌的各种细胞因子在病毒性肝炎转归期起重要作用[3],白细胞介素10（in...丙型肝炎病毒（hepatitis C virus,HCV）是导致慢性肝炎的主要病原之一^（[1]）,HCV感染后20%～30%患者能自主清除,近70%可发展为慢性感染^（[2]）。免疫应答过程中分泌的各种细胞因子在病毒性肝炎转归期起重要作用[3],白细胞介素10（interleukin 10,IL-10）和IL-12在宿主防御、免疫稳态中起重要调节作用。展开更多
Objective To study the effect of gene radiotherapy combining injection of recombinant plasmid pNEgr-mIL-12 with local X-irradiation on cancer growth and to elucidate the mechanisms of tumor inhibition. Methods Alkalin...Objective To study the effect of gene radiotherapy combining injection of recombinant plasmid pNEgr-mIL-12 with local X-irradiation on cancer growth and to elucidate the mechanisms of tumor inhibition. Methods Alkaline lysis was used to extract the plasmid and polyethylene glycol 8000 (PEG 8000) was applied for further purification of plasmids. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of IL-12 protein. C57BL/6J mice were subcutaneously inoculated with B16 melanoma cells and the plasmid was injected directly into the tumor. Gene-radiotherapy combining pNEgr-mIL-12 recombinant plasmid with X-irradiation was given three times to C57BL/6J mice bearing B16 melanoma. Changes in immunologic parameters of tumor-bearing mice were detected with relevant immunologic assays. Results Results showed a significant decrease in tumor growth rate (P<0.05-0.001) after 3 times of gene-radiotherapy with IL-12 and X-irradiation. Immunologic studies showed a significant increase in CTL and NK cytolytic activity (P<0.05-0.001) and an up-regulated secretion of IFN-γ and TNF-α (P<0.01-0.001). Moreover, the expression of mIL-12 in B16 melanoma cells of the treated tumor-bearing mice was found to be higher than that of control. Conclusion pNEgr-mIL-12 plasmid combined with X-irradiation can increase tumor control and the mechanism of increased tumor inhibition is related to the enhancement of anticancer immunity in tumor-bearing mice.展开更多
Objective: To detect the existence of immune tolerance induced by gamma-ray irradiation. Methods: Peritoneal cells were harvested from mice subjected to 5 Gy 60Co gamma-ray total body irradiation at 3d, 7d, 15d and 30...Objective: To detect the existence of immune tolerance induced by gamma-ray irradiation. Methods: Peritoneal cells were harvested from mice subjected to 5 Gy 60Co gamma-ray total body irradiation at 3d, 7d, 15d and 30d, then their counts, morphological changes and IL-12 gene expression were investigated. Results: After irradiation, the peritoneal cells were sharply reduced, the cell morphology shifted from round-like to polymorphic and fusiform with some processes, expression of IL-12 p35 was seriously suppressed, while that of IL-12 p40 greatly enhanced. Conclusion: Our data highly suggest that the gamma-ray irradiation could potentially induce dendritic cell (DC) commitment and immune tolerance.展开更多
To investigate the antitumor activity of IL 12, the induction of differentiation of IL 12 was observed using erythroleukemia cells (FBL 3) as model. After incubation with 200 U/mL IL 12 for 48 h, DNA synthesis of FBL ...To investigate the antitumor activity of IL 12, the induction of differentiation of IL 12 was observed using erythroleukemia cells (FBL 3) as model. After incubation with 200 U/mL IL 12 for 48 h, DNA synthesis of FBL 3 cells in S phase decreased significantly; the expression of CD14 which is the specific marker of monocyte increased, the rate of NBT + cells was apparently higher than that of the untreated FBL 3 cells. After treating FBL 3 cells with IL 12 for 72 h, the expression of 33D1 and NLDC145 which are the specific markers of dendritic cells increased markedly, the surface molecules such as MHC II,B7 1, B7 2, and VCAM 1 were up regulated; morphological observation showed two kinds of cells: some cells had a ruffled surface and plentiful lysosome; the others had many dendritic projections on the surface, and contained numerous mitochondria. Functionally, the IL 12 treated FBL 3 cells could apparently stimulate the proliferation of allogeneic and autologous T lymphocytes, and improve the specific cytotoxic activity of CTL on FBL 3 cells. These results indicated that erythroleukemia cells were induced by IL 12 to differentiate into the monocytes and dendritic cells, then exhibited the antigen presenting function. The data outline a new mechanism for IL 12 to treat leukemia.展开更多
文摘Considering that both innate and adaptive immune responses are involved in the pathogenesis of Crohn's disease(CD), novel therapeutic options have significantly been developed. Biological agents represent an important addition to the conventional treatments for immuno-inflammatory conditions, acting as antagonists of adhesion molecules or various inflammatory cytokines. The interleukin 12(IL-12)/IL-23 common pathway has been found to play a determinant role in the induction of inflammation in adaptive immune responses. In particular, IL-23 promotes the differentiation of na?ve T helper cells into Th17 phenotype with the concomitant secretion of several inflammatory cytokines such as IL-17 and IL-22, whereas IL-12 induces the Th1 polarization and production of critical cytokines such as interferon-γ and tumor necrosis factor. Nowadays, there is increased interest regarding the role of IL-23 as a therapeutic target of CD through the blockage of IL-23 mediated pathways. In this editorial, we focus on the role of IL-12/IL-23 pathway in the regulation of mucosal immunity and in the induction and maintenance of chronic inflammation. In parallel, we critically discuss the available data regarding the therapeutic effect of the IL-12/IL-23 inhibitors and especially of ustekinumab, a human monoclonal antibody which has been recently approved by the United States Food and Drug Administration for the management of moderateto-severe CD and its potential to be used as first-line therapy in everyday clinical practice.
文摘The full-length cDNA encoding the subunits p40 and p35 of human interleukin12(hIL12) were cloned separately by RTPCR, linked together by internal ribosomal entry site (IRES) of encephalomyocarditis virus which initiates capindependent translation to form a dicistronic gene fragment. The dicistronic fragment was placed between the cytomegalovirus (CMV) promoter and SV40 polyA signal to form a dicistronic expression cassette. Subsequently, the dicistronic expression cassette was inserted into E1 region of Ad5 genome in cosmid vector pAx1cw of E1substitution type. By homologous recombination with EcoT22Idigested Ad5 DNATPC in 293 cells, the replicationdeficient recombinant adenoviruses of hIL12 were generated efficiently. After infected with hIL12 recombinant adenoviruses in vitro, 293 cells, human hepatocellular carcinoma cells HepG2, and primary human skin fibroblasts expressed and secreted hIL12 at comparable levels (30～60ng/ 106cells/24hr), which could stimulate the proliferation and IFNγ production of human lymphoblasts. These suggest that the dicistronic adenovirus vector of hIL12 could effectively mediate the expression of bioactive hIL12 and might be used in cancer gene therapy.
文摘丙型肝炎病毒（hepatitis C virus,HCV）是导致慢性肝炎的主要病原之一^（[1]）,HCV感染后20%～30%患者能自主清除,近70%可发展为慢性感染^（[2]）。免疫应答过程中分泌的各种细胞因子在病毒性肝炎转归期起重要作用[3],白细胞介素10（interleukin 10,IL-10）和IL-12在宿主防御、免疫稳态中起重要调节作用。
文摘Objective To study the effect of gene radiotherapy combining injection of recombinant plasmid pNEgr-mIL-12 with local X-irradiation on cancer growth and to elucidate the mechanisms of tumor inhibition. Methods Alkaline lysis was used to extract the plasmid and polyethylene glycol 8000 (PEG 8000) was applied for further purification of plasmids. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of IL-12 protein. C57BL/6J mice were subcutaneously inoculated with B16 melanoma cells and the plasmid was injected directly into the tumor. Gene-radiotherapy combining pNEgr-mIL-12 recombinant plasmid with X-irradiation was given three times to C57BL/6J mice bearing B16 melanoma. Changes in immunologic parameters of tumor-bearing mice were detected with relevant immunologic assays. Results Results showed a significant decrease in tumor growth rate (P<0.05-0.001) after 3 times of gene-radiotherapy with IL-12 and X-irradiation. Immunologic studies showed a significant increase in CTL and NK cytolytic activity (P<0.05-0.001) and an up-regulated secretion of IFN-γ and TNF-α (P<0.01-0.001). Moreover, the expression of mIL-12 in B16 melanoma cells of the treated tumor-bearing mice was found to be higher than that of control. Conclusion pNEgr-mIL-12 plasmid combined with X-irradiation can increase tumor control and the mechanism of increased tumor inhibition is related to the enhancement of anticancer immunity in tumor-bearing mice.
基金This work was supported by the National Natural Science Foundation of China(No. 39670289) and by a grant from Ministry of Education of China.
文摘Objective: To detect the existence of immune tolerance induced by gamma-ray irradiation. Methods: Peritoneal cells were harvested from mice subjected to 5 Gy 60Co gamma-ray total body irradiation at 3d, 7d, 15d and 30d, then their counts, morphological changes and IL-12 gene expression were investigated. Results: After irradiation, the peritoneal cells were sharply reduced, the cell morphology shifted from round-like to polymorphic and fusiform with some processes, expression of IL-12 p35 was seriously suppressed, while that of IL-12 p40 greatly enhanced. Conclusion: Our data highly suggest that the gamma-ray irradiation could potentially induce dendritic cell (DC) commitment and immune tolerance.
文摘To investigate the antitumor activity of IL 12, the induction of differentiation of IL 12 was observed using erythroleukemia cells (FBL 3) as model. After incubation with 200 U/mL IL 12 for 48 h, DNA synthesis of FBL 3 cells in S phase decreased significantly; the expression of CD14 which is the specific marker of monocyte increased, the rate of NBT + cells was apparently higher than that of the untreated FBL 3 cells. After treating FBL 3 cells with IL 12 for 72 h, the expression of 33D1 and NLDC145 which are the specific markers of dendritic cells increased markedly, the surface molecules such as MHC II,B7 1, B7 2, and VCAM 1 were up regulated; morphological observation showed two kinds of cells: some cells had a ruffled surface and plentiful lysosome; the others had many dendritic projections on the surface, and contained numerous mitochondria. Functionally, the IL 12 treated FBL 3 cells could apparently stimulate the proliferation of allogeneic and autologous T lymphocytes, and improve the specific cytotoxic activity of CTL on FBL 3 cells. These results indicated that erythroleukemia cells were induced by IL 12 to differentiate into the monocytes and dendritic cells, then exhibited the antigen presenting function. The data outline a new mechanism for IL 12 to treat leukemia.
