EgM123重组蛋白抗血清对多房棘球绦虫原头蚴体外发育的影响  

Effects of specific antibodies against EgM123 recombinant protein on the development of Echinococcus multilocularis protocercariae in vitro

作　　者：赵莉 王冰洁 班万里 艾尔肯·艾沙 段兰利 陈云英 郭璐 余婉蓉 张壮志 ZHAO Li;WANG Bing-jie;BAN Wan-li;Erkin Eysa;DUAN Lan-li;CHEN Yun-ying;GUO Lu;YU Wan-rong;ZHANG Zhuang-zhi(Veterinary Research Institute,Animal Science Academy of Xinjiang(Animal Clinical Medicine Research Center,Animal Science Academy of Xinjiang),Urumqi,Xinjiang 830013,China;Collage of Veterinary Medicine of Xinjiang Agricultural University;Xinjiang Xingmuyuan Agriculture Limited Liability Com party)

机构地区：[1]新疆畜牧科学院兽医研究所(新疆畜牧科学院动物临床医学研究中心),新疆乌鲁木齐830013 [2]新疆农业大学动物医学学院 [3]新疆兴牧源农业有限责任公司

出　　处：《中国病原生物学杂志》2021年第7期778-782,共5页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.32060802);新疆维吾尔自治区公益性科研院所项目(No.KY2014013)。

摘　　要：目的探讨EgM123重组蛋白抗血清对多房棘球绦虫原头蚴体外发育的影响。方法采集新鲜活力强的多房棘球绦虫原头蚴,体外培养48 h,经1%亚甲基蓝染色计数,原头蚴存活率>90%,将原头蚴按150~200枚/孔加入24孔板。实验分三组:实验组pET41b-EgM123抗血清组和pET28a-EgM123抗血清组以及对照组,实验组设有50、100、200μl 3个不同剂量。按分组加入不同血清,于37℃、5%CO2培养箱中培养3~6 d,期间定时观察原头蚴存活情况,计算原头蚴死亡率。结果体外培养3 d,对照组的原头蚴死亡率为7.74%,pET41b-EgM123组(50、100、200μl)的原头蚴死亡率分别为35.56%、40.67%和62.98%,pET28a-EgM123组(50、100、200μl)的原头蚴死亡率分别为63.9%、85.84%和97.11%;培养6 d,对照组的原头蚴死亡率为7.2%,pET41b-EgM123组(50、100、200μl)的原头蚴死亡率分别为21.55%、26.21%和52.29%,pET28a-EgM123组(50、100、200μl)的原头蚴死亡率分别为80.98%、77.16%和98.64%。pET41b-EgM123和pET28a-EgM123抗血清与对照组相比,对原头蚴杀伤效果显著(P<0.01),pET28a-EgM123抗血清与pET41b-EgM123相比,杀伤效果更显著(P<0.05),抗血清浓度与原头蚴的被损伤程度和死亡率成正比。结论 EgM123抗血清对多房棘球绦虫原头蚴有杀伤作用,以pET28a-EgM123抗血清杀伤效果更显著,可能与抗血清介导的补体经典激活途径有关。Objective To investigate the effects of EgM123 recombinant protein antiserum on the development of Echinococcus multilocularis protoscoleces in vitro. Methods Fresh and vigorous E. multilocularis protoscoleces were collected and cultured in vitro for 48 h. The survival rate of protoscoleces was higher than 90% after 1% methylene blue staining. Protoscoleces were added to 24-well plates, with 150-200 protoscoleces per well. Protoscoleces were divided into three groups: a pET41 b-EgM123 antiserum group, a pET28 a-EgM123 antiserum group, and the control group. The 2 experimental groups were treated with three doses of antiserum: 50, 100, and 200 μl. Different sera were added to each group, and protoscoleces were cultured in an incubator at a temperature of 37℃ in a 5% CO2 atmosphere for 3 to 6 d. During that period, the survival of protoscoleces was observed regularly, and the mortality rate was calculated. Results After in vitro culture for 3 d, protoscolex mortality in the control group was 7.74%, that in the pET41 b-EgM123 group was 35.56%(50 μl), 40.67%(100 μl), and 62.98%(200 μl), and that in the pET28 a-EgM123 group was 63.9%(50 μl), 85.84%(100 μl), and 97.11%(200 μl). After in vitro culture for 6 d, protoscolex mortality in control was 7.2%, that in the pET41 b-EgM123 group was 21.55%(50 μl), 26.21%(100 μl), and 52.29%(200 μl), and that in the pET28 a-EgM123 group was 80.98%(50 μl), 77.16%(100 μl), and 98.64%(200 μl). Compared to the control group, the pET41 b-EgM123 and pET28 a-EgM123 antisera had a marked killing effect on protocercariae(P<0.01), and the killing effect of pET28 a-EgM123 was more significant than that of pET41 b-EgM123(P<0.05). The concentration of antiserum is proportional to the extent of damage to and the mortality of protoscoleces. Conclusion EgM123 antiserum had a killing effect on E. multilocularis protocercariae, and the killing effect of pET28 a-EgM123 antiserum was more marked than that of pET41 b-EgM123. This might be related to the classical complement activation pa

关 键 词：多房棘球绦虫原头蚴 体外培养 免疫杀伤 

分 类 号：R383.3[医药卫生—医学寄生虫学]