重组华支睾吸虫亲环素A的免疫保护作用

Protective effect of immunization with recombinant Clonorchis sinensis CyPA

目的探讨华支睾吸虫亲环素A(CsCyPA)免疫对华支睾吸虫感染及其所致肝损伤的保护作用。方法将10只雄性SD大鼠随机分成两组,实验组进行重组华支睾吸虫亲环素A(rCsCyPA)免疫,对照组注射同等体积的磷酸盐缓冲液(PBS),实验组平均抗体滴度达到1∶20 000后,对实验组和对照组大鼠进行囊蚴灌胃感染(300个囊蚴/只)。40 d后,连续3 d粪检计算减卵率;3个月后处死大鼠,回收成虫计算减虫率,收集血清检测丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶(AST)水平,并取肝脏胆管周围组织进行HE及Masson染色。结果实验组大鼠相对于对照组减虫率及减卵率分别为31.5%和46.2%;实验组大鼠血清中肝脏功能生化指标AST、ALT水平较对照组得到有效控制,而相对于正常大鼠差异无统计学意义(P>0.05);HE及Masson染色显示实验组大鼠肝脏胆管周围腺瘤样病变、炎性细胞浸润、胶原沉积的程度相对对照组明显减轻。结论 rCsCyPA是一个较好的抗感染和抗病理损伤的保护性抗原。

Objective To study the protective effects of immunization with recombinant Clonorchis sinensis Cyclophilin A（CsCyPA）on Clonorchis sinensis infection and hepatic injury induced by clonorchiasis. Methods 10 male SD rats were randomly divided into two groups. The experimental group was immunized with recombinant Clonorchis sinensis Cyclophilin A（rCsCyPA）while the control group was injected with equal volume of phosphate buffer solution（PBS）. Each rat in both groups was given 300 metacercaria of Clonorchis sinensis by gavage when the titer of anti-CsCyPA antibodies in experimental group rats reached 1∶20 000. After 40 days, continuous fecal examinations for 3 days were conducted to detect the egg reduction. Rats were sacrificed after 3 months of infection. The worm reduction was calculated according to the difference in the number of adults in the hepatobiliary tube of two groups. Sersums were separated to detect the levels of aspartate transaminase（AST）and alanine aminotransferase（ALT）, and liver tissues near the bile duct were stained with HE or Masson.Results The worm reduction and the egg reduction of the experimental group were 31.5% and 46.2%, respectively. In addition, comparing to control group, the AST and ALT levels in serum of the experimental group were significantly reduced,while there was no statistically significant difference compared with normal rats. HE and Masson staining showed ameliorative adenomatoid lesions, inflammatory cell infiltration and collagen deposition near the hepatic bile duct in experimental group mice. Conclusion rCsCyPA is a promising protective antigen against Clonorchis sinensis infection and pathological damage induced by clonorchiasis.