BACKGROUND: Lipopolysaccharide-binding protein (LBP) participates in the interaction of lipopolysacchaide (LPS) with CD14 to modulate the expression of cytokines. Human gingival fibroblast may actively participate in LPS-induced immuno-inflammatory responses through CD14, toll-like receptor (TLR) superfamily, MD-2 and related adaptive proteins, leading to the expression of cytokines. OBJECTIVES: The present in vitro study aimed to investigate the possible effect of LBP and E. coli LPS interaction on the expression of cellular LPS receptors and IL-6 by human gingival fibroblast. METHODS: The mRNA expression of CD14, LBP, TLR-2, TLR-4, MD-2 and IL-6 in human gingival fibroblast explants was detected by reverse transcriptionpolymerase chain reaction (RT-PCR) in the presence or absence of E. coli LPS and recombinant human LBP (rhLBP), while IL-6 peptides were analyzed by ELISA and immunohistochemistry, respectively. RESULTS: Human gingival fibroblast could constitutively express CD14, MD-2 and IL-6 mRNAs, but not TLR-2, TLR-4 and LBP mRNAs. E. coli LPS induced the messages expression of MD-2, TLR-2 and -4. The expression of both IL-6 message and peptide was up-regulated by E. coli LPS in a dose dependent manner. Whereas rhLBP could significantly down-regulate the expression of both mRNAs and peptides of CD14 and IL-6 but not MD-2 signals in the presence or absence of E. coli LPS. The up-regulated expression of TLR-2 and -4 by E. coli LPS no longer existed in the presence of rhLBP. CONCLUSIONS: This study suggests that LBP may down-regulate the expression of IL-6 by human gingival fibroblast. Further studies are warranted to clarify the molecular mechanisms of LBP in regulation of cytokine expression by host cells and to elaborate the relevant clinical implications.
BACKGROUND:Lipopolysaccharide-binding protein (LBP) participates in the interaction of lipopolysacchaide (LPS) with CD14 to modulate the expression of cytokines. Human gingival fibroblast may actively participate in LPS-induced immuno-inflammatory responses through CD14, toll-like receptor (TLR) superfamily, MD-2 and related adaptive proteins, leading to the expression of cytokines. OBJECTIVES: The present in vitro study aimed to investigate the possible effect of LBP and E. coli LPS interaction on the expression of cellular LPS receptors and IL-6 by human gingival fibroblast. METHODS: The mRNA expression of CD14, LBP, TLR-2, TLR-4, MD-2 and IL-6 in human gingival fibroblast explants was detected by reverse transcriptionpolymerase chain reaction (RT-PCR) in the presence or absence of E. coli LPS and recombinant humanLBP (rhLBP), while IL-6 peptides were analyzed by ELISA and immunohistochemistry, respectively. RESULTS:Human gingival fibroblast could constitutively express CD14, MD-2 and IL-6 mRNAs, but not TLR-2, TLR-4 and LBP mRNAs. E. coli LPS induced the messages expression of MD-2, TLR-2 and -4. The expression of both IL-6 message and peptide was up-regulated by E. coli LPS in a dose dependent manner. Whereas rhLBP could significantly down-regulate the expression of both mRNAs and peptides of CD14 and IL-6 but not MD-2 signals in the presence or absence of E. coli LPS. The up-regulated expression of TLR-2 and -4 by E. coli LPS no longer existed in the presence of rhLBP. CONCLUSIONS: This study suggests that LBP may down-regulate the expression of IL-6 by human gingival fibroblast. Further studies are warranted to clarify the molecular mechanisms of LBP in regulation of cytokine expression by host cells and to elaborate the relevant clinical implications.
Authors: Thanuja D K Herath; Richard P Darveau; Chaminda J Seneviratne; Cun-Yu Wang; Yu Wang; Lijian Jin Journal: PLoS One Date: 2013-03-12 Impact factor: 3.240