ABSTRACT
No abstract available.
Subject(s)
Aged , Humans , Male , Amylases/blood , Bone Marrow/pathology , Electrophoresis, Agar Gel , Gene Rearrangement , Immunohistochemistry , Isoenzymes/blood , Multiple Myeloma/diagnosis , Plasmacytoma/pathology , Pleural Effusion, Malignant/pathologyABSTRACT
BACKGROUND: Tumor necrosis factor alpha (TNF-alpha) is a pleiotropic cytokine fulfilling a broad variety of immunoregulatory functions. Monocytes and macrophages play a pivotal role in inflammation and immune regulation. NF-kappaB and HIF-1 are known to increase expression of the TNF-alpha gene in a separate way. METHODS: Human monocytic leukemia, U937 cells, were transfected using the standard electroporation method for intracellular expression of NF-kappaB and HIF-1. We performed analysis using the mammalian two-hybrid assay and co-immunoprecipitation assay for detection of protein interaction of both proteins. In addition, chromatin immunoprecipitation analysis was performed for examination of NF-kappaB and HIF-1 binding on the TNF-alpha gene promoter. RESULTS: Here we show that NF-kappaB and HIF-1 cooperatively induced an increase in expression of the TNF-alpha gene dependent on promoter activity by the direct protein interaction of these two transcription factors. Hypoxia signaling induced marked enhancement of the transactivation of TNF-alpha promoter by HIF-1 and NF-kappaB. A tandem NF-kappaB/HIF-1 binding site was identified within the TNF-alpha promoter, which acted as a strong enhancer element. Physical association of the Rel domain of NF-kappaB and the N-TD domain of HIF-1 was required. Hypoxia treatment also resulted in a significant increase in the protein interaction of NF-kappaB and HIF-1 in vivo. Both transcription factors were recruited on the chromatin TNF-alpha promoter dependent on hypoxia stimuli. CONCLUSION: The results of this study indicate that a variety of extracellular signals for activation of TNF-alpha gene expression might converge on the transcriptional regulation through the NF-kappaB/HIF-1 signaling pathway.
Subject(s)
Humans , Hypoxia , Binding Sites , Chromatin , Chromatin Immunoprecipitation , Electroporation , Enhancer Elements, Genetic , Gene Expression , Immunoprecipitation , Inflammation , Leukemia , Macrophages , Monocytes , NF-kappa B , Proteins , Transcription Factors , Transcriptional Activation , Tumor Necrosis Factor-alpha , Two-Hybrid System Techniques , U937 CellsABSTRACT
BACKGROUND: The Di(a) antigen has been detected with a relatively higher incidence among the Korean and Southeast Asian population. A 'Type and Screen' procedure is recommended for efficient transfusion, therefore, we perform antibody screening tests using antibody screening panels containing a Di(a) cell. The purpose of this study was to report on the experience of unexpected antibody screening test including a Di(a) cell in the Korean population. METHODS: We analyzed the results of antibody screening testing and identification performed during the recent 11-year period from January 2002 to December 2012. A commercially available three-cell antigen panel (Diacell I, II, Di(a); DiaMed, Murten, Switzerland) was used for antibody screening. Antibodies were identified using a LISS/Coombs gel card and NaCl/Enzyme card, using the DiaMed-ID system (DiaMed, Murten, Switzerland). RESULTS: The frequency of unexpected antibodies was 1.23% (1,918/156,161); the most frequently detected antibodies were anti-E (292 samples), followed by anti-E,c (127 samples), anti-Le(a) (103 samples), and anti-Di(a) (91 samples). CONCLUSION: Results of this study showed that the most identified unexpected antibodies were clinically significant, and, in particular, anti-Di(a) antibodies are detected frequently in the Korean population. Thus, unexpected antibody screening test including a Di(a) cell is thought to be helpful in Korea for safe transfusion.