CpG Motifs in Porphyromonas gingivalis DNA Stimulate Interleukin-6 Expression in Human Gingival Fibroblasts
CpG Motifs in Porphyromonas gingivalis DNA Stimulate Interleukin-6 Expression in Human Gingival Fibroblasts
复制标题
牙龈卟啉单胞菌 DNA 中的 CpG 基序刺激人牙龈成纤维细胞中白细胞介素 6 的表达
作者：
A. Takeshita;K. Imai;S. Hanazawa
摘要我们认为牙龈卟啉单胞菌DNA可能通过表达炎症细胞因子在牙周病中起毒力因子的作用。细菌DNA以剂量依赖的方式显著刺激人牙龈成纤维细胞产生白细胞介素-6 （IL-6）。DNase能消除刺激作用，而RNase不能。在真核生物dna处理的成纤维细胞中未观察到这种刺激作用。细菌DNA也以剂量和治疗时间依赖的方式刺激细胞中IL-6基因的表达。此外，当DNA被CpG基序甲基化酶甲基化时，刺激效应被消除。有趣的是，含有回文基序GACGTC的30个碱基合成寡核苷酸可以刺激细胞中IL-6基因的表达及其蛋白的产生。此外，合成寡核苷酸诱导的该细胞因子基因的表达被二硫代氨基吡咯烷酸和n -乙酰-l-胱氨酸阻断，它们是转录因子NF-κB的有效抑制剂。凝胶迁移迁移实验显示，在合成寡核苷酸处理的细胞中，NF-κB与其一致序列的结合增加。此外，我们使用特异性抗体来对抗构成NF-κB的p50和p65，我们发现一致的序列结合蛋白是NF-κB。这些结果首次证明了P. gingivalisDNA中的内部CpG基序通过刺激NF-κB来刺激人牙龈成纤维细胞中IL-6的表达。
ABSTRACT We suggest here that Porphyromonas gingivalis DNA may function as a virulence factor in periodontal disease through expression of inflammatory cytokine. The bacterial DNA markedly stimulated in a dose-dependent manner interleukin-6 (IL-6) production by human gingival fibroblasts. The stimulatory action was eliminated by treatment with DNase but not RNase. The stimulatory effect was not observed in the fibroblasts treated with eucaryotic DNAs. The bacterial DNA also stimulated in dose- and treatment time-dependent manners the expression of the IL-6 gene in the cells. In addition, the stimulatory effect was eliminated when the DNA was methylated with CpG motif methylase. Interestingly, a 30-base synthetic oligonucleotide containing the palindromic motif GACGTC could stimulate expression of the IL-6 gene and production of its protein in the cells. Furthermore, the synthetic oligonucleotide-induced expression of this cytokine gene was blocked by pyrrolidine dithiocarbamate andN-acetyl-l-cystine, potent inhibitors of transcriptional factor NF-κB. Gel mobility shift assay showed increased binding of NF-κB to its consensus sequence in the synthetic oligonucleotide-treated cells. Also, using specific antibody against p50 and p65, which compose NF-κB, we showed the consensus sequence-binding proteins to be NF-κB. These results are the first to demonstrate that the internal CpG motifs in P. gingivalisDNA stimulate IL-6 expression in human gingival fibroblasts via stimulation of NF-κB.
细菌 DNA 中的 CpG 基序可快速激活免疫。
DOI：
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发表时间：
1996
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Yi,AK;Klinman,DM;Martin,TL;Matson,S;Krieg,AM
通讯作者：
Krieg,AM
影响因子：
10.5
作者：
FUJITA, T;NOLAN, GP;BALTIMORE, D
通讯作者：
BALTIMORE, D
NF-κ B 样转录因子介导人成纤维细胞中 gro 的 IL-1/TNF-α 诱导。
DOI：
--
发表时间：
1991
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Anisowicz,A;Messineo,M;Lee,SW;Sager,R
通讯作者：
Sager,R