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用于 H7N9 亚单位疫苗开发的来自中国仓鼠卵巢（CHO）稳定细胞系的重组血凝素和 PELC/CpG 联合佐剂。

Recombinant hemagglutinin produced from Chinese Hamster Ovary (CHO) stable cell clones and a PELC/CpG combination adjuvant for H7N9 subunit vaccine development.

机构信息

Institute of Biotechnology, National Tsing Hua University, Hsinchu 30013, Taiwan.

National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan, Taiwan.

出版信息

Vaccine. 2019 Nov 8;37(47):6933-6941. doi: 10.1016/j.vaccine.2019.02.040. Epub 2019 Aug 2.

DOI:10.1016/j.vaccine.2019.02.040
PMID:31383491
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7115541/
Abstract

The novel H7N9 avian influenza A virus has caused human infections in China since 2013; some isolates from the fifth wave of infections have emerged as highly pathogenic avian influenza viruses. Recombinant hemagglutinin proteins of H7N9 viruses can be rapidly and efficiently produced with low-level biocontainment facilities. In this study, recombinant H7 antigen was obtained from engineered stable clones of Chinese Hamster Ovary (CHO) cells for subsequent large-scale production. The stable CHO cell clones were also adapted to grow in serum-free suspension cultures. To improve the immunogenicity of the recombinant H7 antigens, we evaluated the use of a novel combination adjuvant of PELC and CpG (PELC/CpG) to augment the anti-H7N9 immune responses in mice. We compared the effects with other adjuvants such as alum, AddaVax (MF59-like), and several Toll-like receptor ligands such as R848, CpG, and poly (I:C). With the PELC/CpG combination adjuvant, CHO cell-expressed rH7 antigens containing terminally sialylated complex type N-glycans were able to induce high titers of neutralizing antibodies in sera and conferred protection following live virus challenges. These data indicate that the CHO cell-expressed recombinant H7 antigens and a PELC/CpG combination adjuvant can be used for H7N9 subunit vaccine development.

摘要

自 2013 年以来，新型 H7N9 禽流感病毒已在中国引发人类感染；第五波感染的一些分离株已成为高致病性禽流感病毒。具有低水平生物安全防护设施即可快速有效地生产重组血凝素蛋白的 H7N9 病毒。在这项研究中，从中国仓鼠卵巢（CHO）细胞的工程稳定克隆中获得了重组 H7 抗原，用于随后的大规模生产。稳定的 CHO 细胞克隆也适应了无血清悬浮培养的生长。为了提高重组 H7 抗原的免疫原性，我们评估了使用新型 PELC 和 CpG（PELC/CpG）组合佐剂来增强小鼠对 H7N9 的免疫反应。我们将其与其他佐剂（如明矾、AddaVax（MF59 样）和几种 Toll 样受体配体（如 R848、CpG 和 poly（I:C））进行了比较。使用 PELC/CpG 组合佐剂，能够诱导血清中中和抗体的高滴度，并在活病毒攻击后提供保护，CHO 细胞表达的 rH7 抗原含有末端唾液酸化的复合 N-糖。这些数据表明，CHO 细胞表达的重组 H7 抗原和 PELC/CpG 组合佐剂可用于 H7N9 亚单位疫苗的开发。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/808e/7115541/fc62ff7b725e/gr7_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/808e/7115541/d3161333f569/gr1_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/808e/7115541/a34803fd64e4/gr2_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/808e/7115541/e90a25f9f1a7/gr3_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/808e/7115541/d3c5fcd85cf7/gr4_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/808e/7115541/f9ad2a42f705/gr5_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/808e/7115541/fa26b122c82e/gr6_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/808e/7115541/fc62ff7b725e/gr7_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/808e/7115541/d3161333f569/gr1_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/808e/7115541/a34803fd64e4/gr2_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/808e/7115541/e90a25f9f1a7/gr3_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/808e/7115541/d3c5fcd85cf7/gr4_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/808e/7115541/f9ad2a42f705/gr5_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/808e/7115541/fa26b122c82e/gr6_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/808e/7115541/fc62ff7b725e/gr7_lrg.jpg

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本文引用的文献

[1]
CRISPR/Cas9 gene editing for the creation of an MGAT1-deficient CHO cell line to control HIV-1 vaccine glycosylation.

PLoS Biol. 2018-8-29

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PLoS One. 2018-8-2

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Recombinant hemagglutinin proteins formulated in a novel PELC/CpG adjuvant for H7N9 subunit vaccine development.

Antiviral Res. 2017-9-22

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MMWR Morb Mortal Wkly Rep. 2017-9-8

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Epidemiology, Evolution, and Pathogenesis of H7N9 Influenza Viruses in Five Epidemic Waves since 2013 in China.

Trends Microbiol. 2017-7-19

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