Prophylactic evaluation of recombinant extracellular superoxide dismutase of Brugia malayi in jird model
Prophylactic evaluation of recombinant extracellular superoxide dismutase of Brugia malayi in jird model
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DOI：
10.1016/j.vaccine.2008.04.061
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发表时间：
2008-07-04
期刊：
影响因子：
5.5
通讯作者：
Reddy, M. V. R.
中科院分区：
文献类型：
--
作者：
Dabir, Snehal;Dabir, Pankaj;Reddy, M. V. R.
用马来丝虫成虫cDNA文库和当地正常血清进行免疫筛选，筛选出几个血清反应性克隆，其中包括EC-SOD，它含有一个612 bp的插入片段，与其它线虫超氧化物歧化酶（SOD）具有显著的核苷酸和推导的氨基酸序列同源性。已知SOD在保护寄生虫免受宿主活性氧的侵害中起重要作用。B的编码区。克隆马来EC-SOD（BmEC-SOD）基因，在大肠杆菌中表达，镍琼脂糖树脂亲和纯化。表达的重组蛋白的SOD活性的天然聚丙烯酰胺凝胶染色显示，SOD活性被氰化钾和过氧化氢灭活，但不被叠氮化钠，表明Cu/Zn-SOD的存在。rBmEC-SOD蛋白在pH.7.0-11.0范围内具有活性。进一步用长爪沙鼠（Merionesunguiculatus）作为易感宿主，研究了重组EC-SOD抗原对B的免疫保护作用。马来丝虫病体外抗体依赖性细胞毒试验和原位免疫小室试验显示免疫沙鼠对微丝蚴的细胞毒活性分别为33.5%和36%，对感染性幼虫的细胞毒活性分别为42.8%和45.5%。提示rBmEC-SOD抗原可诱导实验动物产生抗微丝蚴和感染性幼虫的部分保护性免疫应答。(C)2008爱思唯尔有限公司保留所有权利。
The immunoscreening of Brugia malayi adult cDNA library with pooled endemic normal sera identified several seroreactive clones including, EC-SOD which contained a 612 bp insert and showed significant nucleotide and deduced amino acid sequence homologies with superoxide dismutase (SOD) of other nematode parasites. The SODs are known to play an important role in the protection of parasite against reactive oxygen species of the host. The coding region of the B. malayi EC-SOD (BmEC-SOD) was cloned and expressed in Escherichia coli followed by affinity purification on nickel agarose resin. Staining of native polyacrylamide gel for SOD activity of the expressed recombinant protein revealed that SOD activity inactivated by potassium cyanide and hydrogen peroxide but not by sodium azide, indicating presence of Cu/Zn-SOD. The rBm EC-SOD protein showed its activity over a broad range of pH.7.0-11.0. Further the immune protective activity of recombinant EC-SOD antigen was evaluated in susceptible host, jirds (gerbils) (Meriones unguiculatus) against B. malayi filarial infection. The immunized jirds showed 33.5% and 36% cytotoxicity against microfilariae and 42.8% and 45.5% cytotoxicity against infective larvae in in vitro antibody dependent cellular cytotoxicity (ADCC) assay and in in situ micropore chamber methods respectively. This study suggests that the rBm EC-SOD antigen could stimulate a partial protective immune response against microfilariae and infective larvae in experimental animals against filarial infection. (C) 2008 Elsevier Ltd. All rights reserved.