M13噬菌体展示技术在鉴定肩突硬蜱唾液中免疫原性蛋白的应用。
Application of M13 phage display for identifying immunogenic proteins from tick (Ixodes scapularis) saliva.
作者信息
Becker Martin, Felsberger André, Frenzel André, Shattuck Wendy M C, Dyer Megan, Kügler Jonas, Zantow Jonas, Mather Thomas N, Hust Michael
机构信息
Institut für Biochemie, Biotechnologie und Bioinformatik, Technische Universität Braunschweig, Spielmannstr.7, 38106, Braunschweig, Germany.
University of Rhode Island, URI Center for Vector-Borne Disease, 231 Woodward Hall, 9 East Alumni Avenue, Suite 7, 02881, Kingston, RI, USA.
出版信息
BMC Biotechnol. 2015 May 30;15:43. doi: 10.1186/s12896-015-0167-3.
BACKGROUND
Ticks act as vectors for a large number of different pathogens, perhaps most notably Borrelia burgdorferi, the causative agent of Lyme disease. The most prominent tick vector in the United States is the blacklegged tick, Ixodes scapularis. Tick bites are of special public health concern since there are no vaccines available against most tick-transmitted pathogens. Based on the observation that certain non-natural host animals such as guinea pigs or humans can develop adaptive immune responses to tick bites, anti-tick vaccination is a potential approach to tackle health risks associated with tick bites.
RESULTS
The aim of this study was to use an oligopeptide phage display strategy to identify immunogenic salivary gland proteins from I. scapularis that are recognized by human immune sera. Oligopeptide libraries were generated from salivary gland mRNA of 18 h fed nymphal I. scapularis. Eight immunogenic oligopeptides were selected using human immune sera. Three selected immunogenic oligopeptides were cloned and produced as recombinant proteins. The immunogenic character of an identified metalloprotease (MP1) was validated with human sera. This enzyme has been described previously and was hypothesized as immunogenic which was confirmed in this study. Interestingly, it also has close homologs in other Ixodes species.
CONCLUSION
An immunogenic protein of I. scapularis was identified by oligopeptide phage display. MP1 is a potential candidate for vaccine development.
背景
蜱作为大量不同病原体的传播媒介，其中最著名的或许是莱姆病的病原体伯氏疏螺旋体。美国最主要的蜱传播媒介是黑脚蜱，即肩突硬蜱。蜱叮咬引发了特殊的公共卫生问题，因为针对大多数蜱传播病原体尚无可用疫苗。基于某些非天然宿主动物（如豚鼠或人类）可对蜱叮咬产生适应性免疫反应这一观察结果，抗蜱疫苗接种是应对与蜱叮咬相关健康风险的一种潜在方法。
结果
本研究的目的是使用寡肽噬菌体展示策略，从肩突硬蜱中鉴定出能被人免疫血清识别的具有免疫原性的唾液腺蛋白。从进食18小时的肩突硬蜱若虫的唾液腺mRNA生成寡肽文库。使用人免疫血清筛选出8种具有免疫原性的寡肽。选取3种具有免疫原性的寡肽进行克隆并制备成重组蛋白。用人体血清验证了一种已鉴定的金属蛋白酶（MP1）的免疫原性特征。这种酶此前已有描述，曾被假设具有免疫原性，本研究证实了这一点。有趣的是，它在其他硬蜱物种中也有密切的同源物。
结论
通过寡肽噬菌体展示鉴定出了肩突硬蜱的一种具有免疫原性的蛋白。MP1是疫苗开发的潜在候选物。
Zotero 插件