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目的研究TRL4-NOD2(T4N2)信号传递增强树突状细胞(dendritic cell,DC)抗结核分枝杆菌(MTB)感染机制,为结核病(tuberculosis,TB)的免疫防治提供参考。方法分别用TLR4配体LPS、NOD2配体MDP和T4N2双配体刺激DC后,ELISA法定量检测1h、6h、12h、24h和48h上清液中IL-6和IL-12。DC经MTB感染再用LPS、MDP以及T4N2双配体刺激,24h后分别收集细胞培养并计数细胞内生长的菌落数,判断细菌生长的抑制率。结果ELISA显示T4N2双信号刺激DC后,细胞因子IL-6在1h、6h、12h、24h和48h浓度分别为(86.4±0.34)pg/ml、(92.2±0.69)pg/ml、(93.6±0.69)pg/ml、(96.0±1.73)pg/ml和(107.8±4.53)pg/ml;IL-12在1h、6h、12h、24h和48h浓度分别为(94.6±4.07)pg/ml、(100.9±4.45)pg/ml、(97.9±2.96)pg/ml、(112.4±9.28)pg/ml和(132.8±1.49)pg/ml。不同时间段T4N2双信号刺激较LPS、MDP刺激显著增多(P<0.05);T4N2双信号活化的DC对MTB生长抑制率显著增强。结论 T4N2信号传递能协同增强DC的活化,活化的DC可抑制MTB的生长。
Objective To investigate the mechanism of dendritic cell (MTB) infection by TRL4-NOD2 (T4N2) signaling in order to provide reference for the immunization of tuberculosis (TB). Methods IL-6 and IL-12 in the supernatants of 1h, 6h, 12h, 24h and 48h were detected by ELISA with TLR4 ligand LPS, NOD2 ligand MDP and T4N2 bidentate ligand respectively. DCs were infected with MTB and then stimulated with LPS, MDP and T4N2 double ligands. After 24 hours, the cell cultures were collected and the number of colonies growing within the cells was counted to determine the inhibition rate of bacterial growth. Results After T4N2 stimulation, the concentrations of cytokines IL-6 at 1h, 6h, 12h, 24h and 48h were (86.2 ± 0.34) pg / ml, (92.2 ± 0.69) pg / ml and (93.6 ± 0.69) (96.0 ± 1.73) pg / ml and (107.8 ± 4.53) pg / ml respectively; the concentrations of IL-12 at 1h, 6h, 12h, 24h and 48h were (94.6 ± 4.07) pg / ml and ± 4.45) pg / ml, (97.9 ± 2.96) pg / ml, (112.4 ± 9.28) pg / ml and (132.8 ± 1.49) pg / ml. Compared with LPS and MDP, the stimulation of T4N2 double-stranded DNA significantly increased at different time points (P <0.05). The inhibitory effect of T4N2 double-signal activated DC on MTB growth was significantly increased. Conclusions T4N2 signaling synergistically enhances DC activation, and activated DCs inhibit the growth of MTB.