J Korean Acad Periodontol.  2009 Jun;39(2):177-184. 10.5051/jkape.2009.39.2.177.

Interleukin-8 production and interleukin-8 mRNA expression induced by lipopolysaccharides from Prevotella intermedia and Prevotella nigrescens in monocyte-derived macrophages

• Kim SJ

• KMID: 1733735
• DOI: http://doi.org/10.5051/jkape.2009.39.2.177

Abstract

PURPOSE: Interleukin-8 (IL-8) is an important mediator of immune and inflammatory reactions and is produced by a variety of different cell types. This study was undertaken to investigate the effects of lipopolysaccharides (LPSs) from Prevotella intermedia and Prevotella nigrescens, the major causes of inflammatory periodontal disease, on the production of IL-8 and the expression of IL-8 mRNA in differentiated THP-1 cells, a human monocytic cell line.
METHODS
LPSs from P. intermedia ATCC 25611 and P. nigrescens ATCC 33563 were prepared by the standard hot phenol-water method. THP-1 cells were incubated in the medium supplemented with phorbol myristate acetate to induce differentiation into macrophage-like cells.
RESULTS
We found that LPS preparations from P. intermedia and P. nigrescens can induce IL-8 mRNA expression and stimulate the release of IL-8 in differentiated THP-1 cells without additional stimuli.
CONCLUSIONS
There are no previous reports of the ability of P. intermedia and P. nigrescens LPS to stimulate the release of IL-8, and the present study clearly shows, for the first time, that LPSs from P. intermedia and P. nigrescens fully induced IL-8 mRNA expression and IL-8 production in differentiated human monocytic cell line THP-1. The ability of P. intermedia and P. nigrescens LPS to promote the production of IL-8 may be important in the pathogenesis of inflammatory periodontal disease.

Keyword

interleukin-8; lipopolysaccharide; Prevotella intermedia; Prevotella nigrescens

MeSH Terms

Cell Line
Humans
Interleukin-8
Lipopolysaccharides
Macrophages
Periodontal Diseases
Phorbols
Prevotella
Prevotella intermedia
Prevotella nigrescens
RNA, Messenger
Tetradecanoylphorbol Acetate
Interleukin-8
Lipopolysaccharides
Phorbols
RNA, Messenger
Tetradecanoylphorbol Acetate

Figure

• 

  Figure 1 Dose response of IL-8 production by differentiated THP-1 cells stimulated with P. intermedia (A) and P. nigrescens LPS (B). S. typhimurium LPS was used as a control. Cells were incubated with increasing concentrations of LPS and supernatants were removed after 24 h and assayed for IL-8. The results are means ± standard deviation of four experiments.

• 

  Figure 2 Dose response (A) and time course (B) of IL-8 mRNA expression in differentiated THP-1 cells stimulated with P. intermedia LPS. See Materials and methods for further details. The PCR bands on a gel photograph in one of two separate experiments yielding similar results are shown. (A) Cells were incubated with different concentrations of P. intermedia LPS for 4 h. (B) Cells were incubated in the presence of 10 µg/mL of P. intermedia LPS for different periods of time.

• 

  Figure 3 Dose response (A) and time course (B) of IL-8 mRNA expression in differentiated THP-1 cells stimulated with P. nigrescens LPS. See Materials and methods for further details. The PCR bands on a gel photograph in one of two separate experiments yielding similar results are shown. (A) Cells were incubated with different concentrations of P. nigrescens LPS for 4 h. (B) Cells were incubated in the presence of 10 µg/mL of P. nigrescens LPS for different periods of time.

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