Sialidase Deficiency in Porphyromonas gingivalis Increases IL-12 Secretion in Stimulated Macrophages Through Regulation of CR3, IncRNA GAS5 and miR-21.
Sialidase Deficiency in Porphyromonas gingivalis Increases IL-12 Secretion in Stimulated Macrophages Through Regulation of CR3, IncRNA GAS5 and miR-21.
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牙龈卟啉单胞菌唾液酸酶缺乏通过调节 CR3、IncRNA GAS5 和 miR-21 增加受刺激巨噬细胞中 IL-12 的分泌。
DOI：
10.3389/fcimb.2018.00100
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发表时间：
2018
影响因子：
5.7
通讯作者：
Li C
中科院分区：
文献类型：
--
作者：
Yang X;Pan Y;Xu X;Tong T;Yu S;Zhao Y;Lin L;Liu J;Zhang D;Li C
牙龈卟啉单胞菌（P. gingivalis）是一种主要的牙周病原体，可诱导免疫反应，引发破坏性炎症过程。在炎症过程中，白细胞介素 - 12（IL - 12）会分泌，其与巨噬细胞对细菌的清除相关。最近研究表明，细菌唾液酸酶会影响细菌表面大分子的合成与修饰，且与细菌和宿主细胞之间的相互作用有关。我们之前在牙龈卟啉单胞菌W83中构建了牙龈卟啉单胞菌唾液酸酶基因突变株（ΔPG0352），并发现ΔPG0352的致病性低于野生型菌株。在本研究中，用牙龈卟啉单胞菌W83、ΔPG0352或PG0352互补菌株（comΔPG0352）刺激分化的U937巨噬细胞。透射电子显微镜显示，牙龈卟啉单胞菌导致巨噬细胞膜完整性受损，细胞内细菌被包裹在内吞泡中。与牙龈卟啉单胞菌W83和comΔPG0352刺激的U937细胞相比，ΔPG0352刺激的U937细胞中IL - 12p35和IL - 12p40基因的表达以及IL - 12p70的水平显著更高。为了解释为什么ΔPG0352在巨噬细胞中诱导产生更多的IL - 12，我们进行了免疫荧光测定、PCR芯片以及基因沉默或过表达实验。免疫荧光测定表明，ΔPG0352诱导巨噬细胞中CR3的表达降低。抑制CR3后，牙龈卟啉单胞菌W83、ΔPG0352或comΔPG0352刺激的巨噬细胞之间IL - 12p70水平无显著差异。PCR芯片实验显示，经实时PCR鉴定，牙龈卟啉单胞菌W83和ΔPG0352刺激的巨噬细胞之间miR - 21和长链非编码RNA GAS5（lncRNA GAS5）表达存在差异。CR3阻断以及lncRNA GAS5基因沉默或过表达的实验结果表明，牙龈卟啉单胞菌W83组和ΔPG0352组之间IL - 12水平的差异与CR3、lncRNA GAS5和miR - 21有关。因此可以得出结论，唾液酸酶缺陷菌株通过减弱CR3激活、减少对lncRNA GAS5的抑制、在巨噬细胞中诱导产生较少的miR - 21和较多的IL - 12，从而更容易被清除。这些结果表明，抑制牙龈卟啉单胞菌中唾液酸酶的活性将促使巨噬细胞快速清除该菌。
Porphyromonas gingivalis (P. gingivalis) is a major periodontal pathogen that can induce an immune response leading to a destructive inflammatory process. During the inflammatory process, interleukin-12 (IL-12) is secreted, correlating with bacterial clearance by macrophages. Bacterial sialidase has recently been shown to influence the synthesis and modification of the macromolecules on its surface, and is associated with the interaction between bacteria and host cells. We have previously constructed a P. gingivalis sialidase gene mutant strain in P. gingivalis W83 (ΔPG0352) and found that ΔPG0352 showed less pathogenicity than the wild-type strain. In this study, U937-differentiated macrophages were stimulated by P. gingivalis W83, ΔPG0352, or PG0352 complemented strain (comΔPG0352). Transmission electron microscopy showed that P. gingivalis caused a loss of membrane integrity in macrophages and the intracellular bacteria were enclosed within endocytic vacuoles. The expression of both IL-12p35 and IL-12p40 genes and the levels of IL-12p70 were significantly higher in U937 stimulated by ΔPG0352 than in those with P. gingivalis W83 and comΔPG0352. In order to explain why ΔPG0352 induced more IL-12 in macrophages, immunofluorescence assays, PCR arrays, and gene silence or overexpression experiments were carried out. Immunofluorescence assays showed that ΔPG0352 induced lower expression of CR3 in macrophages. After CR3 was suppressed, there were no significant differences in the IL-12p70 levels between macrophages stimulated by P. gingivalis W83, ΔPG0352 or comΔPG0352. PCR array experiments showed that miR-21 and lncRNA GAS5 were differentially expressed between macrophages stimulated by P. gingivalis W83 and ΔPG0352, which had been identified by real-time PCR. The results of CR3 blocking and lncRNA GAS5 gene silence or overexpression showed that the difference in IL-12 levels between P. gingivalis W83 and ΔPG0352 groups was associated with CR3, lncRNA GAS5 and miR-21. Thus it can be concluded that the sialidase-deficient strain is more easily cleared by attenuating CR3 activation, reducing the inhibition of lncRNA GAS5, inducing less miR-21 and more IL-12 in macrophages. These results indicate that inhibiting the activity of sialidase in P. gingivalis will cause rapid clearing by macrophages.
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影响因子：
56.9
作者：
Lagos-Quintana, M;Rauhut, R;Tuschl, T
通讯作者：
Tuschl, T
影响因子：
2.8
作者：
Gmiterek, Anna;Klopot, Anna;Olczak, Teresa
通讯作者：
Olczak, Teresa
影响因子：
4.4
作者：
Harokopakis, Evlambia;Albzreh, Mohamad H.;Hajishengallis, George
通讯作者：
Hajishengallis, George
影响因子：
2.1
作者：
Highfield, J.
通讯作者：
Highfield, J.
影响因子：
6.4
作者：
Jia, Ling-Fei;Wei, Su-Bi;Yu, Guang-Yan
通讯作者：
Yu, Guang-Yan