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鉴定用于诊断测试的疥螨抗原蛋白以及可能具有免疫调节作用的非抗原蛋白。

Identification of antigenic Sarcoptes scabiei proteins for use in a diagnostic test and of non-antigenic proteins that may be immunomodulatory.

作者信息

Morgan Marjorie S, Rider S Dean, Arlian Larry G

机构信息

Department of Biological Sciences, Wright State University, Dayton, OH, United States of America.

出版信息

PLoS Negl Trop Dis. 2017 Jun 12;11(6):e0005669. doi: 10.1371/journal.pntd.0005669. eCollection 2017 Jun.

Abstract

BACKGROUND

Scabies, caused by the mite, Sarcoptes scabiei, infects millions of humans, and many wild and domestic mammals. Scabies mites burrow in the lower stratum corneum of the epidermis of the skin and are the source of substances that are antigenic or modulate aspects of the protective response of the host. Ordinary scabies is a difficult disease to diagnose.

OBJECTIVE

The goal of this project was to identify S. scabiei proteins that may be candidate antigens for use in a diagnostic test or may be used by the mite to modulate the host's protective response.

METHODS

An aqueous extract of S. scabiei was separated by 2-dimensional electrophoresis and proteins were identified by mass spectrometry. A parallel immunoblot was probed with serum from patients with ordinary scabies to identify IgM and/or IgG-binding antigens. The genes coding for 23 selected proteins were cloned into E. coli and the expressed recombinant proteins were screened with serum from patients with confirmed ordinary scabies.

RESULTS

We identified 50 different proteins produced by S. scabiei, 34 of which were not previously identified, and determined that 66% were recognized by patient IgM and/or IgG. Fourteen proteins were screened for use in a diagnostic test but none possessed enough sensitivity and specificity to be useful. Six of the 9 proteins selected for the possibility that they may be immunomodulatory were not recognized by antibodies in patient serum.

CONCLUSIONS

Thirty-three proteins that bound IgM and/or IgG from the serum of patients with ordinary scabies were identified. None of the 14 tested were useful for inclusion in a diagnostic test. The identities of 16 proteins that are not recognized as antigens by infected patients were also determined. These could be among the molecules that are responsible for this mite's ability to modulate its host's innate and adaptive immune responses.

摘要

背景

由疥螨(疥螨属)引起的疥疮感染了数百万人类以及许多野生和家养哺乳动物。疥螨在皮肤表皮的角质层下层挖掘洞穴,是抗原性物质或调节宿主保护反应的物质来源。普通疥疮是一种难以诊断的疾病。

目的

本项目的目标是鉴定可能作为诊断测试候选抗原或可能被螨虫用于调节宿主保护反应的疥螨蛋白质。

方法

疥螨的水提取物通过二维电泳分离,蛋白质通过质谱鉴定。用普通疥疮患者的血清检测平行免疫印迹,以鉴定IgM和/或IgG结合抗原。将编码23种选定蛋白质的基因克隆到大肠杆菌中,并用确诊为普通疥疮患者的血清筛选表达的重组蛋白。

结果

我们鉴定出疥螨产生的50种不同蛋白质,其中34种以前未被鉴定,并确定66%的蛋白质被患者的IgM和/或IgG识别。筛选了14种蛋白质用于诊断测试,但没有一种具有足够的敏感性和特异性以有用。为其可能具有免疫调节作用而选择的9种蛋白质中有6种未被患者血清中的抗体识别。

结论

鉴定出33种与普通疥疮患者血清中的IgM和/或IgG结合的蛋白质。测试的14种蛋白质中没有一种可用于诊断测试。还确定了16种未被感染患者识别为抗原的蛋白质的身份。这些可能是负责这种螨虫调节其宿主先天和适应性免疫反应能力的分子。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8fb/5481144/1c319ade0c74/pntd.0005669.g001.jpg

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