Díaz-Zúñiga Jaime, Monasterio Gustavo, Alvarez Carla, Melgar-Rodríguez Samanta, Benítez Alvaro, Ciuchi Pía, García Matías, Arias Jocelyn, Sanz Mariano, Vernal Rolando
Periodontal Biology Laboratory, Department of Conservative Dentistry, Dental School, University of Chile, Santiago, Chile.
J Periodontol. 2015 Jan;86(1):108-19. doi: 10.1902/jop.2014.140326.
Different serotypes of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis have been shown to induce differential dendritic cell (DC) responses. This study investigates whether cytokine and CC-chemokine receptor (CCR) production by DCs stimulated with different serotypes of A. actinomycetemcomitans or P. gingivalis is Toll-like receptor 2 (TLR2) and/or TLR4 dependent.
DCs were obtained from healthy individuals and primed at a multiplicity of infection (MOI) of 10(2) with different A. actinomycetemcomitans or P. gingivalis serotypes in the presence or absence of anti-TLR2 or anti-TLR4 blocking antibodies. TLR2 and TLR4 expression, CCR5 and CCR6 expression, and interleukin (IL)-1β, IL-10, IL-12, and IL-23 expression and secretion were quantified by flow cytometry, real-time reverse-transcription polymerase chain reaction, and enzyme-linked immunosorbent assay.
When DCs were stimulated with serotype b of A. actinomycetemcomitans or serotype K1 of P. gingivalis, higher levels of TLR2 or TLR4, respectively, were detected compared to DCs stimulated with the other serotypes. Similarly, higher levels of cytokines and CCRs were detected in serotype b- or serotype K1-primed DCs compared to the others, and these increased levels positively correlated with levels of TLR2 or TLR4. When TLR2 signaling was blocked using a specific anti-TLR2 monoclonal antibody, serotype b-induced cytokine and CCR expression was inhibited; when TLR4 signaling was blocked, serotype K1-induced response was inhibited.
These results demonstrate that the variability of secretion of cytokines and expression of CCRs detected in DCs stimulated with different serotypes of A. actinomycetemcomitans or P. gingivalis is TLR2 or TLR4 dependent, respectively.
不同血清型的伴放线聚集杆菌和牙龈卟啉单胞菌已被证明可诱导不同的树突状细胞(DC)反应。本研究调查了用不同血清型的伴放线聚集杆菌或牙龈卟啉单胞菌刺激DC产生细胞因子和CC趋化因子受体(CCR)是否依赖Toll样受体2(TLR2)和/或TLR4。
从健康个体获取DC,并在存在或不存在抗TLR2或抗TLR4阻断抗体的情况下,以感染复数(MOI)为10(2) 用不同血清型的伴放线聚集杆菌或牙龈卟啉单胞菌进行致敏。通过流式细胞术、实时逆转录聚合酶链反应和酶联免疫吸附测定法对TLR2和TLR4表达、CCR5和CCR6表达以及白细胞介素(IL)-1β、IL-10、IL-12和IL-23表达及分泌进行定量。
当用伴放线聚集杆菌b血清型或牙龈卟啉单胞菌K1血清型刺激DC时,与用其他血清型刺激的DC相比,分别检测到更高水平的TLR2或TLR4。同样,与其他血清型相比,在b血清型或K1血清型致敏的DC中检测到更高水平的细胞因子和CCR,且这些升高的水平与TLR2或TLR4水平呈正相关。当使用特异性抗TLR2单克隆抗体阻断TLR2信号传导时,b血清型诱导的细胞因子和CCR表达受到抑制;当阻断TLR4信号传导时,K1血清型诱导的反应受到抑制。
这些结果表明,在用不同血清型的伴放线聚集杆菌或牙龈卟啉单胞菌刺激的DC中检测到的细胞因子分泌和CCR表达的变异性分别依赖于TLR2或TLR4。