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目的观察日本血吸虫感染后小鼠肝脏及脾脏自然杀伤(NK)T细胞的动态变化特征。方法将24只6～8周龄雌性BALB/c小鼠随机分为4组,其中3组每鼠经腹部皮肤感染(14±2)条日本血吸虫尾蚴,分别于感染后3、6、12周剖杀小鼠,分离肝脏及脾脏单个核淋巴细胞,应用异硫氰酸荧光素(FITC)标记的CD49b单克隆抗体及藻红蛋白(PE)标记的CD3e单克隆抗体标记细胞,流式细胞仪检测CD3/CD49b双阳性的NKT细胞占单核淋巴细胞的比例。体外刺激实验中将分离的正常小鼠脾脏单个核淋巴细胞,分别用SEA、虫卵蛋白、虫卵脂质作为刺激物,观察其中NKT细胞的比例变化。结果日本血吸虫感染小鼠脾脏中NKT细胞,于感染后12周比例为(4.73±0.41)%,显著高于对照组比例(2.07±0.12)%(P<0.01);肝脏NKT细胞在感染后3周(8.03±0.23)%与对照组(8.60±1.48)%无明显变化,但感染后6周及12周NKT比例明显上升,分别为(15.90±0.76)%和(21.00±0.30)%,与对照组相比差异有统计学意义(P<0.01)。体外实验结果显示,刺激物为SEA和虫卵脂质时,NKT细胞的比例分别为(0.77±0.03)%及(0.80±0.06)%,明显高于空白对照组(0.53±0.03)%及虫卵蛋白组(0.50±0.06)%(P<0.05)。结论日本血吸虫感染导致小鼠脾脏及肝脏中NKT细胞比例上调,其上调可能与虫卵可溶性抗原中的脂质组分有关。
Objective To observe the dynamic changes of natural killer (NK) T cells in mice liver and spleen after Schistosoma japonicum infection. Methods Twenty-four female BALB / c mice aged 6 to 8 weeks were randomly divided into 4 groups. Each group of 3 mice were infected with (14 ± 2) cercariae of Schistosoma japonicum by abdominal skin, Mice were sacrificed and the liver and spleen mononuclear lymphocytes were isolated. FACS-labeled CD49b monoclonal antibody and phycoerythrin (PE) labeled CD3e monoclonal antibody were used to label the cells. Flow cytometry To detect CD3 / CD49b double positive NKT cells accounted for mononuclear lymphocyte ratio. In vitro stimulation experiments will be isolated from normal mice spleen lymphocytes, respectively SEA, egg protein, egg lipids as stimuli, NKT cells were observed in which the proportion of changes. Results The proportion of NKT cells in spleen of Schistosoma japonicum infected mice was (4.73 ± 0.41)% at 12 weeks after infection, significantly higher than that in control group (2.07 ± 0.12)% (P <0.01) (8.03 ± 0.23)% and (8.60 ± 1.48)%, respectively, but the proportion of NKT increased significantly at weeks 6 and 12 after infection (15.90 ± 0.76) and (21.00 ± 0.30)%, respectively The difference between the control group was statistically significant (P <0.01). The results of in vitro experiments showed that the proportion of NKT cells was (0.77 ± 0.03)% and (0.80 ± 0.06)%, respectively, which was significantly higher than that of the blank control group (0.53 ± 0.03)% and the insects Ovalbumin group (0.50 ± 0.06)% (P <0.05). Conclusion Infection with Schistosoma japonicum leads to an up-regulation of NKT cells in spleen and liver of mice, which may be related to lipid components of soluble antigen in eggs.