目的 探讨旋毛虫(Trichinella spiralis)感染小鼠树突状细胞(DC)的C型凝集素-2(dectin-2)的变化情况。 方法 72只雌性BABL/c小鼠随机分成两组,实验组每鼠灌服200条旋毛虫肌幼虫,分别于感染后7、14、21、28、35和42 d各取小鼠6只,分离脾脏和肠系膜淋巴结(MLN),制备单细胞悬液,用流式细胞术检测脾脏和MLN内DC上dectin-2的表达情况;对照组不感染旋毛虫肌幼虫。取C57BL/6小鼠骨髓,体外条件下用粒细胞-巨噬细胞集落刺激因子(GM-CSF)和白细胞介素4(IL-4)诱导培养髓源树突状细胞(BMDC)。培养成熟后加入旋毛虫肌幼虫排泄/分泌抗原(ES)100 μg/ml,同时设PBS对照组,分别于第6、12、24、36和48小时用流式细胞术检测dectin-2的表达量。 结果 流式细胞术检测结果显示,实验组BABL/c小鼠感染旋毛虫肌幼虫后7 d,脾脏中DC的dectin-2表达量为(7.0±0.7)%,对照组为(15.1±1.6)%,实验组明显降低,差异有统计学意义(P<0.01)。实验组小鼠感染旋毛虫肌幼虫后7、21和28 d MLNDC的dectin-2表达量分别为(11.1±3.5)%、(12.4±1.4)%和(6.9±1.0)%,对照组分别为(6.6±0.4)%、(4.9±0.4)%和(4.0±1.4)%,差异有统计学意义(P<0.05)。体外实验结果显示,ES抗原刺激BMDC后,第6、12、24、36、48小时dectin-2的表达量分别为(9.4±2.2)%、(6.9±1.8)%、(6.9±0.7)%、(9.1±1.9)%和(15.9±1.9)%。PBS组为(21.3±6.3)%。第6、12、24、36小时组与PBS组比较表达量降低,差异有统计学意义(P<0.01)。 结论 旋毛虫感染可以引起小鼠树突状细胞上的dectin-2受体表达量发生改变。
Objective To investigate the effect of Trichinella spiralis infection on dectin-2 expression on dendritic cells(DC) in mice. Methods Seventy-two female BABL/c mice were randomized into experimental and control groups(n=36 in each), and received intragastric administration of 200 muscle larvae of Trichinella spiralis and PBS respectively. Six animals were each sacrificed on days 7, 14, 21, 28, 35 and 42 after administration. The spleen and mesenteric lymph nodes(MLN) were dissected and single cell suspensions were prepared. The expression of dectin-2 on DC in the spleen and MLN was examined by flow cytometry. In another experiment, bone marrow was obtained from C57BL/6 mice, and was treated with granulocyte-macrophage colony-stimulating factor and interleukin 4 to induce bone marrow-derived dendritic cells(BMDC) in vitro. Then 100 μg/ml T. spiralis excretory/secretory(ES) antigen or the same volume of PBS was added. The expression of dectin-2 was determined by flow cytometry at 6, 12, 24, 36 and 48 h. Results The flow cytometry results showed a significant decrease of dectin-2 expression[(7.0±0.7)%] on spleen DC compared with that of the control group[(15.1±1.6)%](P〈0.01) on day 7, and significant increase of dectin-2 on MLNDC compared with that of the control on days 7, 21 and 28 [(11.1±3.5)% vs. (6.6±0.4)%, (12.4±1.4)% vs. (4.9±0.4)%, (6.9±1.0)% vs. (4.0±1.4)%] (P〈0.05) after infection. In vitro results showed that dectin-2 expression was (9.4±2.2)%, (6.9±1.8)%, (6.9±0.7)%, (9.1±1.9)%, and(15.9±1.9)% respectively at 6, 12, 24, 36, and 48 h after ES stimulation on BMDC. The expression at 6, 12, 24 and 36 h decreased significantly in comparison to that of the PBS group[(21.3±6.3)%](P〈0.01). Conclusion Trichinella infection can induce changes of dectin-2 expression on DC of mice.