论文部分内容阅读
目的探讨Toll样受体7(TLR7)敲除对日本血吸虫感染小鼠免疫应答的影响。方法野生型C57BL/6小鼠(WT,n=9)和TLR7-/-敲除小鼠(TLR7-/-,n=9)以腹部贴片法感染日本血吸虫尾蚴(20条)。感染后42 d处死小鼠,以胸主动脉灌注法收集虫体(n=6),并取肝脏进行虫卵计数;分别取WT、TLR7-/-未感染小鼠,WT、TLR7-/-感染小鼠的脾脏(n=3),制备脾细胞悬液;加入日本血吸虫虫卵(250个/ml)刺激72 h,ELISA法检测γ干扰素(IFN-γ)、肿瘤坏死因子α(TNF-α)、白细胞介素4(IL-4)和IL-10水平。结果感染后42 d,WT组平均虫荷数为(10.5±3.3)条,每克肝卵数为(38 251.9±4 891.5)个;TLR7-/-组平均虫荷数为(9.8±5.2)条,每克肝卵数为(38 160.9±3 341.0)个,两组间差异无统计学意义(P>0.05)。在未感染的情况下,TLR7-/-小鼠脾细胞上清液中的IL-10[(1702.6±572.3)pg/ml]和IL-4[(59.5±10.1)pg/ml]水平远高于WT组小鼠[(595.2±386.3)和(8.3±0.9)pg/ml](P<0.05,P<0.01)。感染后42 d,TLR7-/-组小鼠脾细胞上清液中的TNF-α[(43.7±9.8)pg/ml]和IFN-γ[(215.2±35.4)pg/ml]水平低于WT组[(63.4±22.9)和(383.5±253.3)pg/ml],而IL-4[(63.9±33.9)pg/ml]水平高于WT组小鼠[(23.3±11.5)pg/ml],但TLR7-/-组与WT组间的差异无统计学意义(P>0.05)。结论 TLR7-/-小鼠在未感染的状态下偏向Th2反应,不影响感染日本血吸虫6周后的免疫应答。
Objective To investigate the effect of Toll-like receptor 7 (TLR7) knockout on immune response in mice infected with Schistosoma japonicum. Methods C57BL / 6 mice (WT, n = 9) and TLR7 - / - mice (TLR7 - / -, n = 9) were infected with cercariae of Schistosoma japonicum (20) by abdominal patch method. Mice were sacrificed at 42 days after infection, and the worms were harvested by thoracic aorta perfusion (n = 6). Liver samples were taken for counting of eggs. WT, TLR7 - / - uninfected WT mice, WT, TLR7 - / - (N = 3) in the spleen of the mice (n = 3) to prepare a suspension of spleen cells; stimulation with Schistosoma japonicum eggs (250 / ml) for 72 h; ELISA for the detection of IFN-γ, -α), interleukin 4 (IL-4) and IL-10 levels. Results The average number of worms in WT group was (10.5 ± 3.3) days after infection, and the number of liver cells per gram was (38 251.9 ± 4 891.5). The mean number of worms in TLR7 - / - group was (9.8 ± 5.2) There was no significant difference between the two groups (P> 0.05). IL-10 [(1702.6 ± 572.3) pg / ml] and IL-4 [(59.5 ± 10.1) pg / ml] in TLR7 - / - mice splenocyte supernatants were much higher (595.2 ± 386.3) and (8.3 ± 0.9) pg / ml] in WT group (P <0.05, P <0.01). At 42 days after infection, the levels of TNF-α [(43.7 ± 9.8) pg / ml] and IFN-γ [(215.2 ± 35.4) pg / ml] in the supernatant of TLR7 - / - (63.4 ± 22.9) and (383.5 ± 253.3) pg / ml, respectively, while the level of IL-4 in 63.9 ± 33.9 pg / ml was significantly higher than that of WT mice [(23.3 ± 11.5) pg / However, there was no significant difference between TLR7 - / - group and WT group (P> 0.05). Conclusion TLR7 - / - mice are biased towards Th2 response in the uninfected state and do not affect the immune response of Schistosoma japonicum after 6 weeks.