ABSTRACT
Objective:To investigate the distribution of γδT17,Th17 and Tc17 cells in the lung of mice severely infected by influenza A(H1N1)pdm09 virus and the relationship between these cells with lung immunopathalogical injury.Methods:Intranasal infection was used to establish mouse model of severe H1N1 infection.Flow cytometry assay was used to detect the proportion and number of γδT17 cells,Th17 cells and Tc17 cells in the lung.The concentrations of interleukin-17A(IL-17A),interleukin-1β(IL-1β)and interleukin-23(IL-23) in the bronchoalveolar lavage fluid and serum were assayed by enzyme-linked immunosorbent assay and Lu-minex assay.Results:①The model of mice severely infected by influenza A(H1N1)pdm09 virus was established successfully.②The ratio of γδT cells,but not CD4+T and CD8+T cells in total lymphocytes of the lung of infected mice significantly increased compared with uninfected control mice at the third day post infection(DPI)(P<0.01).③The proportion and number of γδT17 cells,Th17 cells and Tc17 cells in total γδT cells,Th cells and Tc cells in the lung of infected mice were significant higher than that in uninfected control mice at the first DPI,respectively.However,the absolute number of γδT17 cells was far more than Th17 and Tc17 cells(P<0.05);④The concentration of IL-17A in BALF increased significantly after infection(P<0.05),and the concentration of IL-17A in serum increased significantly at the third DPI(P<0.05).The concentrations of both IL-1β and IL-23 in BALF probably participating in the activation of γδT17 cells increased significantly after infection compared with uninfected control mice.Conclusion:The γδT17 cells could be activated and secreted IL-17A via γδTCR non-depended pathway and involved in inflammatory pathological injury of lung at the early stage of severe H1N1 infection.
ABSTRACT
Objective:To establish an experimental model for intracellular antibacteria and endotoxin neutralization in vitro to detect the antibacterial and endotoxin neutralization activity of the muBPI_(25) protein.Methods: RAW264.7 cells were transfected with pcDNA3.1(+)muBPI_(36-259), and then were infected with intracellular bacterial of either G ~+/G~-to establish the experimental model of intracellalar antibacteria.The RAW264.7 cells were co-transfected with the pSecTag2B-muBPI_(36-259) and dual-luciferase reporter gene plasmids for establishment of the experimental model of endotoxin neutralization.Results:The experimental model of intracellular antibacteria confirmed that the muBPI_(25) protein could inhibit/kill Salmonella typhi.The experimental model of endotoxin neutralization indicated that the muBPI_(25) protein could neutralize endotoxin.Conelusion: We firstly demonstrate that murine BPI N-terminal functional fragment(muBPI_(25) protein)can inhibit/kill Salmonella typhi,and can neutralize, its lysating product, endotoxin.
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Objective To investigate the inhibitive effect of antisense VEGF165 cDNA on growth and angiogensis of human neuroblastoma in nude mouse models.Methods Three kinds of SH-SY5Y cells which had been stably transfected by sense VEGF165 cDNA,antisense VEGF165 cDNA and empty vector respectively,were inoculated subcutaneously to nude mice.The size of subcutaneous tumors was measured,and the morphology of tumor tissue was observed by microscope.The microvessel density(MVD)in tumor mass was analyzed by CD31 immunohistochemistry staining.Apoptotic cells were detected by the electron microscope.Results The growth of tumor mass,which was inoculated with antisense VEGF,significantly decreased in nude mice compared with empty vector group(P
ABSTRACT
Objective The present study was designed to explore the inhibitory effect of antisense VEGF-(165) cDNA on angiogenesis,growth rate of human neuroblastoma.Methods The eukaryotic expression vectors bearing antisense VEGF-(165) cDNA or sense VEGF-(165) cDNA were constructed.The stable cell lines transfected with the sense or antisense VEGF-(165) cDNA were established by using the selective medium containing 400?mg/L of G418.These cell lines were further studied for the exogenous antisense VEGF mRNA expression by RT-PCR and inhibition of expression of endogenous VEGF protein by immunocytochemical staining and ELISA.The proliferation of the transfected cells was analyzed by MTT method.Transfected cells were subcutaneously transplanted into nude mice and the growth of tumor masses was observed and weighted.Results The antisense VEGF was only detected in SH-SY5Y/AsVEGF cells by RT-PCR.The expression of VEGF protein was dramatically declined in the SH-SY5Y/AsVEGF cells compared with the original cells and empty vector transfected cells by immunocytochemical staining and ELISA.There was no effect of antisense VEGF transfection on cell proliferation in vitro.The growth of tumor mass with the cells transfected with antisense VEGF was significantly decreased in nude mice.Conclusion Antisense VEGF cDNA transfection to SH-SY5Y cells can effectively inhibit the expression of endogenous VEGF protein and tumor growth in nude mice.