ABSTRACT
Objective:To study the influences of Mycoplasma penumoniae capsular polysaccharide (CPS) on the phagocytosis and membrane molecules expression of the human peripheral blood mononuclear cells derived dendritic cells by binding to dendritic cell-specific intercellular adhesion molecule-3-grabbing nonintegrin (DC-SIGN),so as to know the effect of CPS on the maturation of dendritic cells.Methods:M.pneumoniae strain was cultivated and CPS was extracted.Human peripheral blood mononuclear cells were separated and induced to dendritic cells,then identified the cells by flow cytometry and observation under the microscope.CPS was used to treat dendritic cells or cells pretreated with DC-SIGN monoclonal antibody,and then FITC-dextran phagocytosis and surface markers CD83,HLA-DR,CD80 and CD86 were detected by flow cytometry.Results:The dendritic cells tended to form colony groups.The positive rate of CD11c molecule in the cultured dendritic cells was about 86.27％.After stimulated by CPS,the FITC-dextran fluorescence mean intake by dendritic cells were increased (P＜0.05),while the cell surface membrane molecules CD83,HLA-DR,CD80 and CD86 were decreased significantly when compared with the PBS treated control cells(P＜0.05).When blocked DC-SIGN with the monoclonal antibody,the FITC-dextran fluorescence mean and membrane molecules expression had no statistical difference with the control cells(P＞0.05).Conclusion:M.pneumoniae CPS can promote the phagocytic function of DC and inhibit the expression of CD83,HLA-DR,CD80 and CD86.
ABSTRACT
<p><b>BACKGROUND</b>Daxx has been identified as a nuclear protein that involves in apoptosis and transcriptional repression. Daxx co-localizes with the promyelocytic leukemia (PML) protein and regulates transcription. Human Daxx (hDaxx) is a protein that functions as a transcriptional regulation through its interaction with some DNA-associated proteins. The aim of this study was to explore the transcriptional regulatory effect of hDaxx interacting with adenovirus (Ad) 12 E1B (Ad12E1B) 55-kDa oncoprotein.</p><p><b>METHODS</b>The co-localization of hDaxx-Ad12E1B or hDaxx-PML protein in the nucleus was observed under a confocal microscope. Interaction of hDaxx and Ad12E1B was analyzed by yeast two-hybrid assay. Direct binding of hDaxx and Ad12E1B was analyzed using coimmunoprecipitation and Western blot in vivo and in vitro. The activity of a luciferase reporter gene, which was regulated by an hDaxx modulated thymidine kinase (TK) promoter, was detected in an automat luminometer.</p><p><b>RESULTS</b>Ad12E1B, which co-localized with hDaxx in the nuclei of G401-CC3 cells, disrupted the co-localization of hDaxx and PML in the PML oncogenic domains (PODs). hDaxx bound directly to Ad12E1B in vivo and in vitro. hDaxx interacted with Ad12E1B along its full length. Ad12E1B enhanced transcriptional repression activity of hDaxx.</p><p><b>CONCLUSION</b>Ad12E1B disrupts the co-localization of hDaxx with PML in PODs and enhances transcriptional repression activity of hDaxx.</p>
Subject(s)
Humans , Adaptor Proteins, Signal Transducing , Adenovirus E1B Proteins , Physiology , Carrier Proteins , Genetics , Cell Line, Tumor , Intracellular Signaling Peptides and Proteins , Neoplasm Proteins , Nuclear Proteins , Genetics , Promyelocytic Leukemia Protein , Repressor Proteins , Physiology , Transcription Factors , Transcription, Genetic , Tumor Suppressor ProteinsABSTRACT
<p><b>BACKGROUND</b>This study was designed to investigate the potential pathogenicity of Mycoplasma penetrans (M. penetrans) and its molecular mechanisms responsible for the induction of iNOS gene expression in mouse macrophages stimulated by lipid-associated membrane proteins (LAMPs) prepared from M. penetrans.</p><p><b>METHODS</b>Mouse macrophages were stimulated with M. penetrans LAMPs to assay the production of nitric oxide (NO). The expression of inducible nitric oxide synthase (iNOS) was detected by RT-PCR and Western blotting. The activity of nuclear factor kappaB (NF-kappaB) and the effects of pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-kappaB, on the production of nitric oxide and the expression of iNOS were also assessed in mouse macrophages treated with M. penetrans LAMPs by indirect immunofluorescence and Western blotting.</p><p><b>RESULTS</b>M. penetrans LAMPs stimulated mouse macrophages to produce nitric oxide in a dose- and time-dependent manner. The mRNA and protein levels of iNOS were also upregulated in response to LAMP stimulation and inhibited by PDTC treatment. M. penetrans LAMPs were found to trigger NF-kappaB activation, a possible mechanism for the induction of iNOS expression.</p><p><b>CONCLUSION</b>This study demonstrated that M. penetrans may be an important etiological factor of certain diseases due to the ability of M. penetrans LAMPs to stimulate the expression of iNOS, which is probably mediated through the activation of NF-kappaB.</p>