日本血吸虫酚氧化酶抗原诱导免疫对病鼠肝脏病理变化的影响  

EFFECT OF PHENOL OXIDASE PROTEIN ANTIGEN ON LIVER PATHOLOGIC CHANGE IN MICE INFECTED WITH SCHISTOSOMA JAPONICUM

作　　者：章莹[1] 蔡国斌[2] 蒋明森[2] 何立[2] 杨孟祥[2] 

机构地区：[1]武汉大学医学院医学遗传学教研室,武汉430071 [2]武汉大学医学院人体寄生虫学教研室,血吸虫病研究室

出　　处：《中国血吸虫病防治杂志》2004年第4期246-248,F002,共4页Chinese Journal of Schistosomiasis Control

基　　金：国家自然科学基金专项基金项目 ( No.3 0 2 40 0 74);湖北省教育厅资助项目 ( No.2 0 0 1A14 0 0 9)

摘　　要：目的　观察日本血吸虫酚氧化酶天然蛋白抗原诱导免疫对病鼠肝脏的病理变化。方法　将 4 2 d活成虫置于含 0 .0 5 %戊巴比妥钠的 RPMI 16 4 0培养基中孵育 8h后洗净 ,匀浆 ,超声粉碎、低温高速离心 ,将含酚氧化酶的上清液进行聚丙烯酰胺凝胶电泳 ,嗣后分别在两侧切取胶宽约 1cm进行酶染色 ,解剖刀准确切下相应的未染色的凝胶。冻干后碾成粉末 ,高速电动匀浆成浆糊状 ,冷浸过夜。高速离心得上清即为日本血吸虫酚氧化酶粗抗原。设立 3组进行免疫实验 :实验组、佐剂对照组和空白对照组。初次免疫后第 2、4周 ,各重复 1次加强免疫 ,最后 1次免疫 10 d后用新鲜逸出尾蚴 (4 0± 1)条 /鼠进行攻击感染。 4 2 d剖杀动物 ,观察日本血吸虫感染小鼠肝脏的病理变化。结果　与对照组相比 ,酚氧化酶免疫实验组小鼠肝脏表面较光滑 ,隐约可见少量灰白色小点 ,肝色泽略带灰色 ,质地较软 ;压片观察小鼠肝脏内虫卵集聚数量明显减少 ,且以未成熟虫卵为主。肝脏内伴少量肝细胞坏死 ,细胞浸润不明显 ,可见较多未成熟卵周围无肉芽肿反应。小鼠肝组织内虫卵肉芽肿的平均直径和面积比对照组小鼠肝组织内虫卵肉芽肿的平均直径和面积明显减小 ,两组间差异有非常显著性 (P<0 .0 1)。结论　日本血吸虫酚氧化酶蛋白具抗原性 。Objective To explore the effect of phenol oxidase antigen on liver pathologic change in mice infected with Schistosoma japonicum. Methods 42d-aged adult worms were incubated in RPMI 1640 containing 0.05% sodium phenobarbital for 8 h. The worms were washed three times with PBS (pH 6.8) and homogenized with a Teflon pestle. The homogenate was then centrifuged at 3000 g for 20 min at 4 ℃. Supernatant fractions containing phenol oxidase (PO) were analyzed by means of polyacrylamide gel electrophoresis (PAGE). The rude antigen of PO was obtained by cutting the corresponding gel of PO activities. Three groups were set up to observe whether PO could induce protective immunity: experiment group, adjuvant control group and water control group. On day 42 post infection with (40±1) cercariae of Schistosoma japonicum, the mice were sacrificed to observe liver pathologic changes. Results The liver surface of PO immunized group was rather smooth and the liver color was slightly gray. A few pale nods were seen indistinctly but not clearly. Necrosis and inflammatory cell infiltration were not clear. There were many immature eggs without granuloma reaction. The mean diameter and area of the granuloma in the experiment group were less than those in the control group. There were significant differences among the 3 groups (P<0.01). Conclusion Phenol oxidase protein has antigenicity and it can induce protective immunity of host's resisting schistosomiasis.

关 键 词：日本血吸虫 酚氧化酶 肝脏病理变化 

分 类 号：R383.24[医药卫生—医学寄生虫学]