PRODUCTION OF SINGLE CHAIN FRAGMENT VARIABLE (scFv) ANTIBODY AGAINST PARANOSEMA LOCUSTAE ALPHA/BETA-HYDROLASE AND IMMUNOLOCALIZATION OF MICROSPORIDIAN ENZYME IN THE INFECTED HOST CELL.

Q4 Medicine

Tsitologiya Pub Date : 2017-01-01

V V Dolgikh, A A Tsarev, I V Senderskiy, S A Timofeev

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引用次数: 0

Abstract

Microsporidia is a widespread group of fungi-related intracellular parasites. Direct contact of the most microsporidia species with host cytoplasm suggests that these parasites may control physiological processes of infected cells by secretion of various proteins. In previous experiments, secretion of significant amounts of microsporidia Paranosema locustae alpha/beta-hydrolase into infected cells of Locusta migratoria fat bodies was demonstrated using polyclonal antibodies against the enzyme. However, heterologous expression of microsporidian hydrolase in yeast Pichia pastoris cells was not accompanied by its secretion. In this study, we have constructed library of recombinant single chain antibodies (scFv-fragments) against proteins of fat bodies of infected locusts and isolated mini-antibody specifically recognizing the studied enzyme using phage display technology. Immunoblotting and immunofluorescent microscopy with selected scFv-fragment confirmed secretion of two different in size forms of P. locustae alpha/beta-hydrolase into infected host cell. Prospects of scFv-fragment use to explore the role of microsporidian hydrolase in host-parasite relations and mechanism of its secretion are discussed in the paper.

本刊更多论文

单链片段可变抗体(scFv)的产生和微孢子虫酶在被感染宿主细胞中的免疫定位。

微孢子虫是一种分布广泛的与真菌相关的细胞内寄生虫。大多数微孢子虫与宿主细胞质的直接接触表明，这些寄生虫可能通过分泌各种蛋白质来控制被感染细胞的生理过程。在先前的实验中，使用针对该酶的多克隆抗体证明了向迁移蝗虫脂肪体感染细胞分泌大量的副病微孢子虫α / β水解酶。然而，微孢子虫水解酶在酵母毕赤酵母细胞中的异源表达不伴有分泌。在本研究中，我们构建了针对感染蝗虫脂肪体蛋白的重组单链抗体(scFv-fragments)文库，并利用噬菌体展示技术分离出特异性识别所研究酶的微抗体。免疫印迹和免疫荧光显微镜与选定的scfv片段证实了两种不同大小形式的P.蝗虫α / β -水解酶分泌到感染的宿主细胞。本文对利用scfv片段探索微孢子虫水解酶在宿主-寄生虫关系中的作用及其分泌机制进行了展望。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Tsitologiya Medicine-Pathology and Forensic Medicine

CiteScore

0.40

自引率

0.00%

发文量

期刊介绍： TSITOLOGIYA publishes papers on all chief problems of cell biology: morphology, physiology, immunology, biochemistry, molecular biology, biophysics. The submitted manuscripts may be in Russian or in English. The Journal accepts the original papers not published previously and dealing with studies on both animal and plant cells, reviews, discussional papers, communications on new methods of investigations, reviews of books published the current year, chronicle. Communications on research meetings (congresses, conferences, symposia, etc.) for the Chronicle Section of the Journal are accepted only if submitted not later than 2 months after the date of the meeting.