Elevated mRNA expression for supervillin and vascular endothelial growth factor in human neutrophils stimulated with lipopolysaccharide from Porphyromonas gingivalis.
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.
Morozumi
T
T
Department of Periodontology, Faculty of Dentistry, Niigata University, Japan.
Kubota
T
T
Sugita
N
N
Ohsawa
Y
Y
Yamazaki
K
K
Yoshie
H
H
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
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