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新型流感 HA 三聚体蛋白的结构与应用，用于评估 HA 茎特异性免疫。

Structure and applications of novel influenza HA tri-stalk protein for evaluation of HA stem-specific immunity.

机构信息

Department of Infection and Immunity, Luxembourg Institute of Health, Esch-sur-Alzette, Luxembourg.

Latvian Biomedical Research and Study Centre, Riga, Latvia.

出版信息

PLoS One. 2018 Sep 27;13(9):e0204776. doi: 10.1371/journal.pone.0204776. eCollection 2018.

DOI:10.1371/journal.pone.0204776
PMID:30261065
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6160157/
Abstract

Long alpha helix (LAH) from influenza virus hemagglutinin (HA) stem or stalk domain is one of the most conserved influenza virus antigens. Expression of N-terminally extended LAH in E. coli leads to assembly of α-h elical homotrimer which is structurally nearly identical to the corresponding region of post-fusion form of native HA. This novel tri-stalk protein was able to differentiate between group 1 and 2 influenza in ELISA with virus-infected mice sera. It was also successfully applied for enzyme-linked immunospot assay to estimate the number of HA stem-reactive antibody (Ab)-secreting cells in mice. An in-house indirect ELISA was developed using a HA tri-stalk protein as a coating antigen for evaluation of HA stem-specific Ab levels in human sera collected in Luxembourg from 211 persons with occupational exposure to swine before the pandemic H1N1/09 virus had spread to Western Europe. Our results show that 70% of these pre-pandemic sera are positive for HA stem-specific Abs. In addition, levels of HA stem-specific Abs have positive correlation with the corresponding IgG titers and neutralizing activities against pandemic H1N1/09 virus.

摘要

流感病毒血凝素（HA）茎或柄区的长α螺旋（LAH）是最保守的流感病毒抗原之一。在大肠杆菌中表达 N 端延伸的 LAH 会导致α螺旋三聚体的组装，其结构与天然 HA 融合后形式的相应区域几乎相同。这种新型三柄蛋白能够在 ELISA 中区分组 1 和 2 流感病毒，使用感染病毒的小鼠血清。它还成功地应用于酶联免疫斑点测定法来估计小鼠中 HA 茎反应性抗体（Ab）分泌细胞的数量。使用 HA 三柄蛋白作为包被抗原，开发了一种内部间接 ELISA，用于评估在大流行 H1N1/09 病毒传播到西欧之前，在卢森堡从 211 名接触猪的职业人群中收集的人血清中的 HA 茎特异性 Ab 水平。我们的结果表明，这些大流行前血清中有 70%对 HA 茎特异性 Abs 呈阳性。此外，HA 茎特异性 Abs 的水平与针对大流行 H1N1/09 病毒的相应 IgG 滴度和中和活性呈正相关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eecc/6160157/bb82c2009904/pone.0204776.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eecc/6160157/5d2653223012/pone.0204776.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eecc/6160157/2f9e093164dc/pone.0204776.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eecc/6160157/cb35d145562f/pone.0204776.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eecc/6160157/627cd9d7c561/pone.0204776.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eecc/6160157/bb82c2009904/pone.0204776.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eecc/6160157/5d2653223012/pone.0204776.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eecc/6160157/2f9e093164dc/pone.0204776.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eecc/6160157/cb35d145562f/pone.0204776.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eecc/6160157/627cd9d7c561/pone.0204776.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eecc/6160157/bb82c2009904/pone.0204776.g005.jpg

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