Zhang Chi, Xu Chenrong, Gao Li, Li Xiting, Zhao Chuanjiang
Department of Periodontology, Hospital of Stomatology, Sun Yat-sen University, Guangzhou, China.
Guangdong Provincial Key Laboratory of Stomatology, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou, China.
PeerJ. 2021 Apr 23;9:e11094. doi: 10.7717/peerj.11094. eCollection 2021.
To investigate the effect and mechanism of () lipopolysaccharide (LPS) on Th17 cell differentiation mediated by CD14 monocytes.
. LPS-activated CD14 monocytes were co-cultured with CD4T cells in different cell ratios. An indirect co-culture system was also established using transwell chambers. Furthermore, anti- Delta-like ligand 4 (Dll-4) antibody was used to investigate the role of Dll-4 in Th17 cell response. The mRNA expression was analyzed using quantitative reverse transcription-polymerase chain reaction, and secreted cytokines in culture supernatant were detected using enzyme-linked immunosorbent assay. Flow cytometry was used to determine the frequencies of Th17 cells. IL-17 protein expression levels were determined using western blotting assay.
LPS increased the expressions of interleukin (IL)-1, IL-6, IL-23 and transforming growth factor (TGF)- in CD14 monocytes. Th17 cell frequency upregulated, which is not solely cytokine-dependent but rather requires cell-cell contact with activated monocytes, particularly in the 1:10 cell ratio. Furthermore, . LPS increased t he expression of Dll-4 on CD14 monocytes, whereas the anti- Dll-4 a ntibody decreased the response of Th17 cells. The results suggest that . LPS enhances Th17 cell response via Dll-4 upregulation on CD14 monocytes.
研究（）脂多糖（LPS）对CD14单核细胞介导的Th17细胞分化的影响及机制。
......将LPS激活的CD14单核细胞与CD4 T细胞以不同细胞比例共培养。还使用Transwell小室建立间接共培养系统。此外，使用抗Delta样配体4（Dll-4）抗体研究Dll-4在Th17细胞反应中的作用。使用定量逆转录-聚合酶链反应分析mRNA表达，并使用酶联免疫吸附测定法检测培养上清液中的分泌细胞因子。流式细胞术用于确定Th17细胞的频率。使用蛋白质印迹法测定IL-17蛋白表达水平。
LPS增加了CD14单核细胞中白细胞介素（IL）-1、IL-6、IL-23和转化生长因子（TGF）-的表达。Th17细胞频率上调，这不仅依赖于细胞因子，还需要与活化的单核细胞进行细胞间接触，特别是在1:10的细胞比例下。此外，......LPS增加了CD14单核细胞上Dll-4的表达，而抗Dll-4抗体降低了Th17细胞的反应。结果表明，......LPS通过上调CD14单核细胞上的Dll-4增强Th17细胞反应。
