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哺乳动物细胞稳定表达牛暂时热病毒分泌 GΔTM 蛋白的免疫原性。

The immunogenicity of the secretory GΔTM protein of bovine ephemeral fever virus stably expressed by mammalian cells.

机构信息

Interdisciplinary Graduate Program in Genetic Engineering, The Graduate School, Kasetsart University, Bangkok, 10900, Thailand.

Center for Advance Studies in Agriculture and Food, KU Institute Studies, Kasetsart University, Bangkok, 10900, Thailand.

出版信息

Vet Microbiol. 2019 Jun;233:113-117. doi: 10.1016/j.vetmic.2019.04.032. Epub 2019 Apr 28.

Abstract

Bovine ephemeral fever virus (BEFV) causes an acute febrile disease in cattle and water buffalo. The disease has an impact on dairy and beef production in tropical and subtropical countries. Vaccination is used for disease prevention and control. In this study, we developed a recombinant lentivirus to produce mammalian stable cells expressing histidine-tagged BEFV G protein with a deleted transmembrane domain (GΔTM) as a secretory protein. In addition, guinea pigs were immunised with the purified GΔTM protein and booster immunised at a 3-week interval. The mammalian stable cells were able to continuously produce GΔTM protein for a minimum of 25 passages. All of the mammalian stable cells expressing GΔTM protein could react specifically with a BEFV convalescent bovine serum. Serum samples from the immunised guinea pigs could react strongly and specifically with the purified GΔTM protein. Moreover, post-immunised guinea pig sera contained antibodies that could neutralise BEFV. These results indicate that the G protein without a transmembrane domain can be used as a subunit vaccine for the prevention and control of BEFV. The availability of the mammalian stable cells, which constitutively express GΔTM protein, could facilitate the potential use of the secretory protein for BEFV diagnosis and vaccine development.

摘要

牛暂时热病毒(BEFV)可引起牛和水牛的急性发热疾病。该病对热带和亚热带国家的奶牛和肉牛生产有影响。疫苗接种用于疾病的预防和控制。在本研究中,我们开发了一种重组慢病毒,用于生产表达组氨酸标记的 BEFV G 蛋白的哺乳动物稳定细胞,该 G 蛋白缺失跨膜域(GΔTM),作为一种分泌蛋白。此外,豚鼠用纯化的 GΔTM 蛋白免疫,并在 3 周间隔进行加强免疫。哺乳动物稳定细胞能够连续产生 GΔTM 蛋白至少 25 代。所有表达 GΔTM 蛋白的哺乳动物稳定细胞都能与 BEFV 恢复期牛血清特异性反应。免疫豚鼠的血清样本能与纯化的 GΔTM 蛋白强烈且特异性反应。此外,免疫后的豚鼠血清中含有能中和 BEFV 的抗体。这些结果表明,无跨膜域的 G 蛋白可用作 BEFV 的预防和控制的亚单位疫苗。表达 GΔTM 蛋白的哺乳动物稳定细胞的可用性,可能促进该分泌蛋白在 BEFV 诊断和疫苗开发中的潜在应用。

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