Toll-like receptor-2 and-4 responses regulate neutrophil infiltration into the junctional epithelium and significantly contribute to the composition of the oral microbiota
Toll-like receptor-2 and-4 responses regulate neutrophil infiltration into the junctional epithelium and significantly contribute to the composition of the oral microbiota
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DOI：
10.1002/jper.18-0719
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发表时间：
2019-10-01
影响因子：
4.3
通讯作者：
Darveau, Richard P.
中科院分区：
文献类型：
--
作者：
Chang, Ana M.;Liu, Quanhui;Darveau, Richard P.
背景口腔牙龈组织，尤其是结合上皮（JE），经常暴露于龈下菌斑。牙龈健康的一个关键组成部分是调节中性粒细胞的数量，这些中性粒细胞迁移到牙龈缝隙中以抵消其有害影响。本报告调查的贡献，先天防御受体，Toll样受体（TLR）2，TLR 4，和两者（TLR 2/4）的中性粒细胞的稳态维持在乙脑。方法使用靶向16 S核糖体RNA基因V3-V4区的MiSeq平台，从12- 14周龄TLR 2、TLR 4、TLR 2/4双敲除（KO）小鼠收集的全口腔拭子中分析细菌组成。对下颌骨进行组织学检查，以定量JE中的中性粒细胞和骨丢失。最后，使用定量实时PCR定量总细菌负荷。结果与野生型小鼠相比，TLR基因敲除小鼠JE中的中性粒细胞募集明显增加（P = 0.0079）。此外，TLR 4和TLR 2/4 KO小鼠的细菌数量显著增加（分别为P = 0.0022和P = 0.0152）。最后，口腔微生物组的比较组成分析显示，每种KO菌株都具有独特的微生物群落，这些微生物群落彼此不同，但保持了相似的牙槽骨水平。结论正常小鼠乙脑中神经元的迁移不需要TLR 2或TLR 4。然而，TLR 2和TLR 4 KO小鼠中嗜中性粒细胞数量的显著增加以及口腔微生物组成的显著变化表明，这些TLR有助于健康小鼠牙周组织中细菌和宿主之间的稳态关系。
Background Oral gingival tissue, especially the junctional epithelium (JE), is constantly exposed to sub-gingival plaque. A key component of gingival health is the regulation of the number of neutrophils that migrate into the gingival crevice to counteract its harmful effects. This report investigates the contribution of innate defense receptors, Toll-like receptor (TLR)2, TLR4, and both (TLR2/4) to the maintenance of neutrophil homeostasis in the JE. Methods Bacterial composition was analyzed from whole oral swabs collected from 12- to 14-week-old TLR2, TLR4, TLR2/4 double knock-out (KO) mice using a MiSeq platform targeting the V3-V4 region of the 16S ribosomal RNA gene. Mandibles were histologically examined for quantification of neutrophils in the JE and bone loss. Lastly, total bacterial load was quantitated using quantitative real-time PCR. Results Compared with wild-type, all TLR KO mice displayed significantly increased recruitment of neutrophils (P = 0.0079) into the JE. In addition, TLR4 and TLR2/4 KO mice demonstrated a significant increase in the number of bacteria (P = 0.0022 and P = 0.0152, respectively). Lastly, comparative compositional analyses of the oral microbiome revealed that each KO strain harbored unique microbial communities that are distinct from each other but maintained similar levels of alveolar bone. Conclusions Neutrophil migration into healthy mouse JE does not require TLR2 or TLR4. However, a significant increase in the number of neutrophils as well as a significant change in the oral microbial composition in both TLR2 and TLR4 KO mice demonstrate that these TLRs contribute to the homeostatic relationship between bacteria and the host in healthy mice periodontal tissue.