Document Detail

cDNA representational difference analysis of human neutrophils stimulated by GM-CSF.
MedLine Citation:
PMID:  11032736     Owner:  NLM     Status:  MEDLINE    
Neutrophils are the first cell type to migrate out of the vascular space and into the inflammatory site during an acute inflammation. However, in chronic inflammatory diseases, such as chronic obstructive pulmonary disease (COPD), a lack of clearance of neutrophils, imbalance between inflammatory mediators produced by neutrophils and their natural inhibitors make these cells a potential cause of tissue destruction in lung disease. Neutrophilic inflammation is generally characterised by high levels of local expression of activating cytokines (e.g., GM-CSF). Only a few studies have been published so far that have investigated the expression of genes preferentially expressed in activated neutrophils. The isolation of such genes, however, can lead to a better understanding of inflammatory disease and the identification of potential novel therapeutic targets or markers of the disease. We performed representational difference analysis of cDNA, a sensitive PCR-based subtractive enrichment procedure, and isolated 12 genes, 1 EST clone, and 3 sequences not represented in the public databases. Differential expression for 9 of these clones was confirmed by Northern hybridisation. Of the above nine transcripts three were chosen and shown to be up-regulated in neutrophils cocultured with stimulated primary human bronchial epithelial cells using a semiquantitative RT-PCR approach. Among the known genes identified were HM-74, CIS1, Cathepsin C, alpha-enolase, CD44, and the gene Translocation Three Four (TTF), most of them previously not known to be involved in GM-CSF induced neutrophil activation. Along with its tissue and cellular distribution we also derived the complete cDNA sequence and genomic structure of CIS1 using an in silico approach. In addition, we also report the initial characterisation of a novel gene, P1-89 that is primarily expressed in granulocytes and is up-regulated in activated cells. Our results identify several important genes associated with neutrophil activation and can lead to a better understanding of the molecular mechanisms of neutrophilic inflammations.
S Yousefi; P R Cooper; B Mueck; S L Potter; G Jarai
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Biochemical and biophysical research communications     Volume:  277     ISSN:  0006-291X     ISO Abbreviation:  Biochem. Biophys. Res. Commun.     Publication Date:  2000 Oct 
Date Detail:
Created Date:  2000-11-06     Completed Date:  2000-11-30     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  0372516     Medline TA:  Biochem Biophys Res Commun     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  401-9     Citation Subset:  IM    
Copyright Information:
Copyright 2000 Academic Press.
Novartis Horsham Research Centre, Wimblehurst Road, Horsham, West Sussex, RH12 5AB, United Kingdom.
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MeSH Terms
Amino Acid Sequence
Antigens, CD44 / genetics
Base Sequence
Blotting, Northern
Bronchi / metabolism
Cathepsin C / genetics
Cells, Cultured
Coculture Techniques
DNA, Complementary / metabolism*
Epithelial Cells / metabolism
Expressed Sequence Tags
Gene Library
Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology*
Granulocytes / metabolism
Immediate-Early Proteins / genetics
Models, Genetic
Molecular Sequence Data
Neutrophils / metabolism*
Phosphopyruvate Hydratase / biosynthesis,  genetics
Proteins / genetics*
RNA, Messenger / metabolism
Reverse Transcriptase Polymerase Chain Reaction
Sequence Analysis, DNA
Sequence Homology, Nucleic Acid
Suppressor of Cytokine Signaling Proteins
Tissue Distribution
Transcription Factors
rho GTP-Binding Proteins
Reg. No./Substance:
0/Antigens, CD44; 0/DNA, Complementary; 0/Immediate-Early Proteins; 0/Proteins; 0/RNA, Messenger; 0/RhoH protein, human; 0/Suppressor of Cytokine Signaling Proteins; 0/Transcription Factors; 0/cytokine inducible SH2-containing protein; 83869-56-1/Granulocyte-Macrophage Colony-Stimulating Factor; EC C; EC GTP-Binding Proteins; EC Hydratase

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