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• W2371235036 abstract "Objective: To construct the recombinant adenovirus vector which co-express human IL-17R and FcγRⅡb gene and further identify its characteristics. This study also explores the effect of this recombinant adenovirus vector on the morbidity of asthma during pregnancy,while blocking the signal regulation of IL-17R-FcγRⅡb. Methods: The splicing fragment of IL-17R and FcγRⅡb gene were amplified and sub-cloned to pAdTrack-CMV shuttle plasmid and then was homologously recombined with backbone plasmid pAdeasy-1. After digested and linearized,the recombinant was transfected into AD293 cells with liposome and identified with GFP expression and PCR. Myeloid dendritic cells( DC) were isolated from asthmatic mice during pregnancy and cultured in vitro. The infection efficiency of exogenous genes,carried by recombinant adenovirus,was detected in target cells by fluorescence microscopy. The mRNA transcription level of IL-17R-FcγRⅡb gene,and the expression level of its protein,was determined by RT-PCR and Western blot. The effect of this recombinant adenovirus on the surface markers of the DC,its role to stimulate the proliferation of T cells,and its impact on the secretion of IFN-γ,IL-4 and IL-17,was analyzed by Flow cytometry( FCM),CCK-8 method and ELISA method.Results: It was confirmed that the sequence of our vectors and insert fragments were completely correct. The expression of GFP and the PCR results confirmed that our recombinant adenovirus,carrying the target gene,was successfully packaged. We obtained CD11c DC and the infection efficiency of the adenovirus was 85%. At the mRNA and protein levels,the positive transcription and expression of IL-17R-FcγRⅡb can be amplified in line with expectations. No significant difference was funded in the expression of positive CD11c cells among all the three cell groups. Compared with that in the control group and in the EGFP-DCs group,the expression CD80marker increased in IL-17R-FcγRⅡb-DCs group,while the expression of CD86marker decreased( P 0. 05). In IL-17R-FcγR Ⅱ b-DCs group,the ability of DC cells to stimulate the proliferation of T cell is much lower than the DC cells in the other two groups at the same concentration ratio( P 0. 05). In the supernatant of co-cultured transfection vectors,the levels of IL-4 and IL-17( Th2 / Th17-related cytokine) decreased,the level of IFN-γ( Th1-related cytokine) increased( P 0. 05),while the ratio of IFN-γ / IL-4 / IL-17 increased respectively. Conclusion: IL-17R-FcγRⅡb fusion gene can induce immune tolerance state of DC,inhibit the proliferation of antigenspecific T cells and affect the differentiation of Th1 / Th2 / Th17 subsets by blocking CD86/ CD28co-stimulatory pathway in vitro. It may play an important role on the formation of immune tolerance defects of asthma during pregnancy,so as on the mitigation of airway inflammation's reactions." @default.
• W2371235036 created "2016-06-24" @default.
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• W2371235036 date "2013-01-01" @default.
• W2371235036 modified "2023-09-26" @default.
• W2371235036 title "Effect of IL-17R-FcγRIIb gene on the expression of dendritic cells in vitro and on the activation of lymphocyte's proliferation" @default.
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