title: Elevated mRNA expression for supervillin and vascular endothelial growth factor in human neutrophils stimulated with lipopolysaccharide from Porphyromonas gingivalis.
abstract: Differential gene expression was examined in human neutrophils stimulated with lipopolysaccharide from Porphyromonas gingivalis (P. gingivalis-LPS) using RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR). LPS from Escherichia coli (E. coli-LPS) was used as control. More than 200 differently expressed transcripts were found in 8 subjects showing differential regulation at the transcriptional level. Densitometric analyses revealed that 42-100 genes were upregulated, while 53-116 genes were downregulated by P. gingivalis-LPS compared with E. coli-LPS. The results of sequencing identification showed the presence of supervillin (SVIL) and vascular endothelial growth factor (VEGF) genes in the clones which were upregulated by P. gingivalis-LPS. Consequently, semiquantitative analyses proved that the level of mRNA for SVIL and VEGF were significantly higher in P. gingivalis-LPS-stimulated neutrophils than in other bacterial (E. coli, Actinobacillus actinomycetemcomitans, Prevotella intermedia) LPS- or synthetic lipid A-stimulated neutrophils. Our findings suggest that an elevated mRNA expression for SVIL could be associated with impaired function of neutrophils when stimulated by P. gingivalis-LPS. Further, the VEGF mRNA over-expression might be related to the pathogenesis of P. gingivalis-associated periodontitis. The RAP-PCR technique used in this study enabled us to identify a number of P. gingivalis-LPS regulated genes which hitherto have not been reported.
authors.affiliation: Department of Periodontology, Faculty of Dentistry, Niigata University, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Morozumi
authors.forename: T
authors.initials: T
authors.lastname: Kubota
authors.forename: N
authors.initials: N
authors.lastname: Sugita
authors.forename: Y
authors.initials: Y
authors.lastname: Ohsawa
authors.forename: K
authors.initials: K
authors.lastname: Yamazaki
authors.forename: H
authors.initials: H
authors.lastname: Yoshie
mesh_terms: D000328:Adult; D016976:Aggregatibacter actinomycetemcomitans / Q000502:physiology; D016172:DNA Fingerprinting; D015536:Down-Regulation / Q000235:genetics; D016228:Endothelial Growth Factors / Q000235:genetics*; D004926:Escherichia coli / Q000502:physiology; D005786:Gene Expression Regulation; D006801:Humans; D008050:Lipid A / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008222:Lymphokines / Q000235:genetics*; D008297:Male; D008565:Membrane Proteins / Q000235:genetics*; D008840:Microfilament Proteins / Q000235:genetics*; D009504:Neutrophils / Q000378:metabolism*; D010518:Periodontitis / Q000235:genetics / Q000382:microbiology; D016966:Porphyromonas gingivalis / Q000502:physiology*; D018713:Prevotella intermedia / Q000502:physiology; D020033:Protein Isoforms / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D018709:Statistics, Nonparametric; D014158:Transcription, Genetic; D015854:Up-Regulation / Q000235:genetics; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
chemical_list: D016228:Endothelial Growth Factors; D008050:Lipid A; D008070:Lipopolysaccharides; D008222:Lymphokines; D008565:Membrane Proteins; D008840:Microfilament Proteins; D020033:Protein Isoforms; D012333:RNA, Messenger; C109413:SVIL protein, human; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors