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无血清 HEK293A 细胞培养系统中柯萨奇病毒 A10 的增殖和免疫特性。

Propagation and immunological characterization of coxsackievirus A10 in a serum-free HEK293A cell culture system.

机构信息

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

Institute of Oral Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan.

出版信息

Virus Res. 2023 May;329:199101. doi: 10.1016/j.virusres.2023.199101. Epub 2023 Mar 24.

Abstract

Coxsackievirus A10 (CVA10) is one of enteroviral pathogens that cause the hand, foot, and mouth disease (HFMD). Since CVA10 was reported to be not easily propagated in the Vero cell culture, a feasible manufacture process for producing formalin-inactivated CVA10 vaccine is urgently needed. Several cell lines that commonly used for viral vaccine production was tested for CVA10 (M2014 strain) culture in this study, and our result showed that CVA10 could be easily propagated in the HEK293A cells. A serum-free HEK293A cell culture system was developed for CVA10 production and the yields have reached over 10 TCID/mL. The biochemical and immunogenic properties of CVA10 particles obtained from this serum-free HEK293A culture were identical to our previous study. Two major particles of CVA10 were separated by ultracentrifugation, and only the infectious mature particles were capable of inducing CVA10 neutralizing antibody responses in the mouse immunogenicity studies. Additionally, we found that coxsackievirus A6 and enterovirus A71 could also be easily propagated using this serum-free HEK293A cell culture system. Our results provide a solution to overcome the obstacle in the propagation of CVA10 and facilitate the development of multivalent vaccines for prevention of HFMD.

摘要

柯萨奇病毒 A10(CVA10)是引起手足口病(HFMD)的肠道病毒病原体之一。由于 CVA10 不易在 Vero 细胞培养物中繁殖,因此迫切需要开发一种可行的生产甲醛灭活 CVA10 疫苗的方法。本研究测试了几种常用于病毒疫苗生产的细胞系对 CVA10(M2014 株)的培养,结果表明 CVA10 可在 HEK293A 细胞中容易繁殖。开发了无血清 HEK293A 细胞培养系统用于 CVA10 的生产,产量已超过 10 TCID/mL。从无血清 HEK293A 培养物中获得的 CVA10 颗粒的生化和免疫原性与我们之前的研究相同。通过超速离心分离出 CVA10 的两种主要颗粒,只有传染性成熟颗粒才能在小鼠免疫原性研究中诱导出 CVA10 中和抗体反应。此外,我们发现使用这种无血清 HEK293A 细胞培养系统也可以很容易地繁殖柯萨奇病毒 A6 和肠道病毒 A71。我们的研究结果为克服 CVA10 的繁殖障碍提供了一种解决方案,并有助于开发用于预防 HFMD 的多价疫苗。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e558/10194310/7e08ab509fc0/gr1.jpg

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