Suppr超能文献

糖依赖的重组可溶性三聚血凝素的免疫原性。

Glycan-dependent immunogenicity of recombinant soluble trimeric hemagglutinin.

机构信息

Virology Division, Department of Infectious Diseases and Immunology, Utrecht University, Utrecht, The Netherlands.

出版信息

J Virol. 2012 Nov;86(21):11735-44. doi: 10.1128/JVI.01084-12. Epub 2012 Aug 22.

Abstract

Recombinant soluble trimeric influenza A virus (IAV) hemagglutinin (sHA(3)) has proven an effective vaccine antigen against IAV. Here, we investigate to what extent the glycosylation status of the sHA(3) glycoprotein affects its immunogenicity. Different glycosylation forms of subtype H5 trimeric HA protein (sH5(3)) were produced by expression in insect cells and different mammalian cells in the absence and presence of inhibitors of N-glycan-modifying enzymes or by enzymatic removal of the oligosaccharides. The following sH5(3) preparations were evaluated: (i) HA proteins carrying complex glycans produced in HEK293T cells; (ii) HA proteins carrying Man(9)GlcNAc(2) moieties, expressed in HEK293T cells treated with kifunensine; (iii) HA proteins containing Man(5)GlcNAc(2) moieties derived from HEK293S GnTI(-) cells; (iv) insect cell-produced HA proteins carrying paucimannosidic N-glycans; and (v) HEK293S GnTI(-) cell-produced HA proteins treated with endoglycosidase H, thus carrying side chains composed of only a single N-acetylglucosamine each. The different HA glycosylation states were confirmed by comparative electrophoretic analysis and by mass spectrometric analysis of released glycans. The immunogenicity of the HA preparations was studied in chickens and mice. The results demonstrate that HA proteins carrying terminal mannose moieties induce significantly lower hemagglutination inhibition antibody titers than HA proteins carrying complex glycans or single N-acetylglucosamine side chains. However, the glycosylation state of the HA proteins did not affect the breadth of the antibody response as measured by an HA1 antigen microarray. We conclude that the glycosylation state of recombinant antigens is a factor of significant importance when developing glycoprotein-based vaccines, such as recombinant HA proteins.

摘要

重组可溶性三聚体流感病毒(IAV)血凝素(sHA(3))已被证明是一种有效的 IAV 疫苗抗原。在这里,我们研究了 sHA(3)糖蛋白的糖基化状态对其免疫原性的影响程度。通过在昆虫细胞和不同的哺乳动物细胞中表达,在不存在和存在 N-糖基化修饰酶抑制剂的情况下,或通过酶法去除寡糖,产生了不同糖基化形式的 H5 三体重组 HA 蛋白(sH5(3))。评估了以下 sH5(3)制剂:(i)在 HEK293T 细胞中产生的带有复杂聚糖的 HA 蛋白;(ii)用 kifunensine 处理的 HEK293T 细胞中表达的带有 Man(9)GlcNAc(2) 部分的 HA 蛋白;(iii)来自 HEK293S GnTI(-) 细胞的含有 Man(5)GlcNAc(2) 部分的 HA 蛋白;(iv)昆虫细胞产生的带有低聚糖 N-糖基化的 HA 蛋白;和(v)用内切糖苷酶 H 处理的 HEK293S GnTI(-) 细胞产生的 HA 蛋白,从而携带每个侧链仅由一个 N-乙酰葡萄糖胺组成。不同的 HA 糖基化状态通过比较电泳分析和释放糖的质谱分析得到证实。在鸡和小鼠中研究了 HA 制剂的免疫原性。结果表明,与携带复杂聚糖或单个 N-乙酰葡萄糖胺侧链的 HA 蛋白相比,携带末端甘露糖部分的 HA 蛋白诱导的血凝抑制抗体滴度显著降低。然而,HA 蛋白的糖基化状态并不影响通过 HA1 抗原微阵列测量的抗体反应的广度。我们得出结论,在开发糖蛋白疫苗(如重组 HA 蛋白)时,重组抗原的糖基化状态是一个非常重要的因素。

相似文献

1
Glycan-dependent immunogenicity of recombinant soluble trimeric hemagglutinin.
J Virol. 2012 Nov;86(21):11735-44. doi: 10.1128/JVI.01084-12. Epub 2012 Aug 22.
2
The influenza A virus hemagglutinin glycosylation state affects receptor-binding specificity.
Virology. 2010 Jul 20;403(1):17-25. doi: 10.1016/j.virol.2010.03.047. Epub 2010 May 2.
3
Influenza Virus Hemagglutinin Glycoproteins with Different N-Glycan Patterns Activate Dendritic Cells In Vitro.
J Virol. 2016 Jun 10;90(13):6085-6096. doi: 10.1128/JVI.00452-16. Print 2016 Jul 1.
9
Recombinant influenza H7 hemagglutinins induce lower neutralizing antibody titers in mice than do seasonal hemagglutinins.
Influenza Other Respir Viruses. 2014 Nov;8(6):628-35. doi: 10.1111/irv.12285. Epub 2014 Sep 12.
10

引用本文的文献

3
A plant-produced SARS-CoV-2 spike protein elicits heterologous immunity in hamsters.
Front Plant Sci. 2023 Mar 7;14:1146234. doi: 10.3389/fpls.2023.1146234. eCollection 2023.
4
Establishment of a Pseudovirus Platform for Neuraminidase Inhibiting Antibody Analysis.
Int J Mol Sci. 2023 Jan 25;24(3):2376. doi: 10.3390/ijms24032376.
8
Multimerization- and glycosylation-dependent receptor binding of SARS-CoV-2 spike proteins.
PLoS Pathog. 2021 Feb 8;17(2):e1009282. doi: 10.1371/journal.ppat.1009282. eCollection 2021 Feb.
10
Impact of Protein Glycosylation on the Design of Viral Vaccines.
Adv Biochem Eng Biotechnol. 2021;175:319-354. doi: 10.1007/10_2020_132.

本文引用的文献

1
Profiling of humoral immune responses to influenza viruses by using protein microarray.
Clin Microbiol Infect. 2012 Aug;18(8):797-807. doi: 10.1111/j.1469-0691.2011.03701.x. Epub 2011 Dec 23.
2
Fitness costs limit influenza A virus hemagglutinin glycosylation as an immune evasion strategy.
Proc Natl Acad Sci U S A. 2011 Dec 20;108(51):E1417-22. doi: 10.1073/pnas.1108754108. Epub 2011 Nov 21.
3
A potent and broad neutralizing antibody recognizes and penetrates the HIV glycan shield.
Science. 2011 Nov 25;334(6059):1097-103. doi: 10.1126/science.1213256. Epub 2011 Oct 13.
4
Towards universal influenza vaccines?
Philos Trans R Soc Lond B Biol Sci. 2011 Oct 12;366(1579):2766-73. doi: 10.1098/rstb.2011.0102.
7
Glycosylation focuses sequence variation in the influenza A virus H1 hemagglutinin globular domain.
PLoS Pathog. 2010 Nov 24;6(11):e1001211. doi: 10.1371/journal.ppat.1001211.
8
Enhanced immunogenicity of stabilized trimeric soluble influenza hemagglutinin.
PLoS One. 2010 Sep 1;5(9):e12466. doi: 10.1371/journal.pone.0012466.

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验