J Korean Acad Periodontol.  2004 Jun;34(2):461-474. 10.5051/jkape.2004.34.2.461.

Chemical and Immunobiological Characterization of Lipopolysaccharides from Prevotella intermedia and Prevotella nigrescens

• Kim SJ

• KMID: 2045167
• DOI: http://doi.org/10.5051/jkape.2004.34.2.461

Abstract

The purpose of this study was to assess some biological activities of lipopolysaccharides (LPSs) from P. intermedia and P. nigrescens. LPS was prepared by the standard hot phenol-water method. NO production was assayed by measuring the accumulation of nitrite in culture supernatants. TNF-alpha production was determined by enzyme-linked immunosorbent assay. Western blot analysis of iNOS and analysis of reverse transcription (RT)-PCR products were carried out. LPS from P. intermedia demonstrated higher KDO content than those from two stains of P. nigrescens. LPSs from P. intermedia and P. nigrescens were mitogenic for spleen cells of BALB/C mouse. The present study clearly shows that LPSs from P. intermedia and P. nigrescens fully induced iNOS expression and NO production in RAW264.7 cells in the absence of other stimuli. Moreover, LPSs from P. intermedia and P. nigrescens clearly induced TNF-alpha production in RAW264.7 cells. The biological activities of LPS from P. intermedia was found to be comparable to those of P. nigrescens LPS. The ability of LPSs from P. intermedia and P. nigrescens to promote the production of NO and TNF-alpha may be important in the pathogenesis of inflammatory periodontal disease.

Keyword

Prevotella intermedia; Prevotella nigrescens; Lipopoplysaccharide; B-cell mitogenicity; Nitric oxide; iNOS; TNF-alpha

MeSH Terms

Animals
Blotting, Western
Coloring Agents
Enzyme-Linked Immunosorbent Assay
Lipopolysaccharides*
Mice
Nitric Oxide
Periodontal Diseases
Prevotella intermedia*
Prevotella nigrescens*
Prevotella*
Reverse Transcription
Spleen
Tumor Necrosis Factor-alpha
Coloring Agents
Lipopolysaccharides
Nitric Oxide
Tumor Necrosis Factor-alpha