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布氏冈比亚丝虫 DEAD 盒 RNA 解旋酶的结构建模研究和免疫预防潜力。

Structural modelling studies and immunoprophylactic potential of Brugia malayi DEAD Box RNA helicase.

机构信息

Division of Parasitology, CSIR-Central Drug Research Institute, BS10/1, Sector 10, Jankipuram extension, Sitapur Road, Lucknow 226021, Uttar Pradesh, India.

出版信息

Parasitology. 2013 Jul;140(8):1016-25. doi: 10.1017/S0031182013000322.

Abstract

DEAD Box RNA helicases are essential enzymes that are involved in RNA metabolic processes such as transcription, pre-mRNA splicing, translation initiation and RNA decay. We have previously over-expressed and biochemically characterized an immunodominant cDNA clone encoding DEAD box RNA helicase (BmL3-Helicase) isolated by immunoscreening of the larval stage cDNA library of Brugia malayi. In the current study, the 3D structure was determined and the immunoprophylactic efficacy of BmL3-Helicase was investigated by immunizing Mastomys coucha with the recombinant protein and subsequently challenging with B. malayi infective larvae. The immunization had an adverse outcome on the establishment of challenged larvae resulting in a 67.4% reduction in adult parasite recovery, a 86.7% decrease in the microfilarial density and profound sterility of the recovered female worms. The immune response thus generated was investigated by measuring the levels of specific antibodies including IgG subclasses, reactive oxygen species and cytokines.

摘要

DEAD 盒 RNA 解旋酶是参与 RNA 代谢过程的必需酶,如转录、前体 mRNA 剪接、翻译起始和 RNA 降解。我们之前已经过表达和生化表征了一种免疫显性 cDNA 克隆,该克隆编码通过免疫筛选马来丝虫幼虫 cDNA 文库分离的 DEAD 盒 RNA 解旋酶(BmL3-解旋酶)。在当前的研究中,通过用重组蛋白免疫 Mastomys coucha 并随后用感染性幼虫挑战,确定了 3D 结构,并研究了 BmL3-解旋酶的免疫预防效果。免疫接种对挑战幼虫的建立产生了不利影响,导致成虫寄生虫回收率降低 67.4%,微丝蚴密度降低 86.7%,并且回收的雌性蠕虫具有明显的不育性。通过测量包括 IgG 亚类、活性氧和细胞因子在内的特异性抗体的水平来研究产生的免疫反应。

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