DNA免疫和蛋白质免疫制备抗狂犬病病毒核蛋白IgY的比较。

IF 1.8 4区 农林科学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE
Journal of Poultry Science Pub Date : 2023-06-17 eCollection Date: 2023-01-01 DOI:10.2141/jpsa.2023014
Nanase Kubo, Chun-Ho Park, Satoshi Inoue, Hajime Hatta
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引用次数: 0

摘要

用病毒抗原免疫蛋鸡可以有效地从蛋黄中产生大量的病毒特异性IgY抗体。世界各地都需要提供实用和经济的狂犬病病毒抗体。我们用狂犬病病毒的抗原基因DNA免疫母鸡,从蛋黄中纯化特异性IgY抗体,并对免疫蛋白化学进行表征,以用于诊断。为了通过DNA免疫制备针对狂犬病病毒核蛋白(RV-N)的特异性IgY抗体,用λ-卡拉胶或弗氏完全佐剂预注射蛋鸡以提高局部免疫活性(预免疫刺激),然后用RV-N重组质粒DNA免疫。从免疫母鸡的蛋黄中制备RV-N特异性IgY抗体。为了进行比较,还使用常规蛋白质抗原免疫来诱导RV-N特异性IgY抗体的产生。用RV-N蛋白抗原免疫蛋鸡,并从蛋黄中纯化RV-N特异性IgY。使用通过DNA(具有免疫前刺激)和蛋白质免疫制备的IgY样品检测针对RV-N抗原的结合活性。免疫组织化学染色显示,通过蛋白质免疫制备的IgY抗体在感染病毒的狗的脑切片中强烈检测到病毒抗原,而通过DNA免疫制备IgY抗体则没有。使用市售狂犬病疫苗(灭活病毒)进行酶联免疫吸附测定,该疫苗用10%福尔马林处理并加热(60°C,30分钟和90°C,5分钟)。DNA免疫制备的IgY与变性抗原的反应性较弱,抗原浓度低于蛋白质免疫制备。这些结果表明,有必要开发一种DNA免疫方法来诱导针对与天然和变性抗原强结合的狂犬病病毒的IgY抗体,以制备可用于临床试验中抗原检测的特异性IgY。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Comparison of Anti-rabies Virus Nucleoprotein IgY Prepared by DNA Immunization and Protein Immunization.

Immunization of egg-laying hens with viral antigens efficiently produces large amounts of virus-specific IgY antibodies from egg yolks. A supply of practical and economical antibodies against the rabies virus is being desired worldwide. We immunized hens with the antigen gene DNA of the rabies virus, purified specific IgY antibodies from the egg yolk, and characterized the immuno-protein chemistry for use as a diagnosis. To prepare specific IgY antibodies against rabies virus nucleoprotein (RV-N) by DNA immunization, laying hens were pre-injected with λ-carrageenan or Freund's complete adjuvant to increase local immune activity (pre-immune stimulation), and then immunized with RV-N recombinant plasmid DNA. RV-N-specific IgY antibodies were prepared from egg yolks of immunized hens. For comparison, conventional protein antigen immunization was also used to induce the production of RV-N-specific IgY antibodies. Laying hens were immunized with an RV-N protein antigen and RV-N-specific IgY was purified from egg yolks. The binding activity against RV-N antigens was examined using IgY samples prepared by DNA (with pre-immune stimulation) and protein immunization. Immunohistochemical staining showed that IgY antibodies prepared by protein immunization strongly detected viral antigens in the brain sections of dogs infected with the virus, whereas IgY antibodies prepared by DNA immunization did not. Enzyme-linked immunosorbent assay was performed using a commercially available rabies vaccine (inactivated virus) treated with 10% formalin and heating (60°C, 30 min and 90°C, 5 min). IgY prepared by DNA immunization had weaker reactivity with denatured antigens and lower antigen concentrations than IgY prepared by protein immunization. These results suggest that it is necessary to develop a DNA immunization method for inducing IgY antibodies against the rabies virus that strongly bind to native and denatured antigens to prepare specific IgYs that can be used for antigen detection in clinical tests.

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来源期刊
Journal of Poultry Science
Journal of Poultry Science AGRICULTURE, DAIRY & ANIMAL SCIENCE-
CiteScore
2.80
自引率
13.30%
发文量
26
审稿时长
12 months
期刊介绍: The Journal of Poultry Science will publish original reports and reviews which either make an original contribution to fundamental science or are of obvious application to the industry. Subjects which are covered include: breeding and genetics, nutrition and feeds, physiology, reproduction, immunology, behavior, environmental science, management and housing welfare, processing and products, and health in poultry. Submission of original articles to the Journal is open to all poultry researchers. The review articles are invited papers written by international outstanding researchers. Articles will be published in English, American style.
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