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使用在永生昆虫细胞系中减毒的黄头病毒(YHV)对虾进行疫苗接种的尝试失败。

Failed shrimp vaccination attempt with yellow head virus (YHV) attenuated in an immortal insect cell line.

作者信息

Gangnonngiw Warachin, Kanthong Nipaporn

机构信息

Centex Shrimp, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok 10400, Thailand.

National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Klong 1, Klong Luang, Pratum Thani 12120, Thailand.

出版信息

Fish Shellfish Immunol Rep. 2023 Jan 7;4:100084. doi: 10.1016/j.fsirep.2023.100084. eCollection 2023 Dec.

Abstract

This short paper on yellow head virus Type-1 (YHV-1) of shrimp describes preliminary research on the potential for using YHV-1 attenuated in insect cells to protect shrimp against yellow head disease (YHD). YHV-1 can cause severe mortality in the cultivated shrimp  and .  No practical vaccination has been reported. The C6/36 mosquito cell cultures inoculated with YHV-1 become positive by PCR and by immunocytochemistry (immunopositive) for up to 30 split-cell passages. Shrimp injected with homogenates from low-passage cultures die from typical YHV-1 disease while shrimp injected with homogenates from high passage cultures do not, even though they become PCR positive and immunopositive for YHV-1. This suggested that viral attenuation had occurred during insect-cell passaging, and it opened the possibility of using homogenates from high-passage insect cultures as a vaccine against YHV-1. To test this hypothesis, homogenates from 30th-passage, YHV-positive cultures were injected into shrimp followed by challenge with virulent YHV-1. Controls were injected with homogenate from 30th-passage, naive (normal stock) insect-cell cultures. No shrimp mortality occurred following injection of either homogenate, but shrimp injected with the YHV-1 homogenate became both RT-PCR positive and immunopositive. Upon challenge 10 days later with YHV-1, mortality in shrimp injected with naive insect-cell homogenate was 100% within 7 days post-challenge while 100% mortality in the YHV-1 homogenate group did not occur until day 9 post-challenge. Kaplan-Meier log-rank survival analysis revealed that survival curves for the two groups were significantly different ( < 0.001). The cause of delay in mortality may be worthy of further investigation.

摘要

这篇关于虾类黄头病毒1型(YHV-1)的短文描述了利用在昆虫细胞中减毒的YHV-1保护虾类免受黄头病(YHD)侵害的潜力的初步研究。YHV-1可导致养殖虾类严重死亡,且尚未有实际可行的疫苗接种报道。接种YHV-1的C6/36蚊细胞培养物在多达30次传代培养中通过PCR检测呈阳性且免疫细胞化学检测呈阳性(免疫阳性)。注射低代培养物匀浆的虾死于典型的YHV-1疾病,而注射高代培养物匀浆的虾虽对YHV-1呈PCR阳性和免疫阳性,但却未死亡。这表明在昆虫细胞传代过程中病毒发生了减毒,这为使用高代昆虫培养物匀浆作为抗YHV-1疫苗开辟了可能性。为验证这一假设,将第30代YHV阳性培养物的匀浆注射到虾中,随后用强毒YHV-1进行攻毒。对照组注射第30代未感染(正常细胞株)昆虫细胞培养物的匀浆。注射两种匀浆后均未出现虾类死亡,但注射YHV-1匀浆的虾RT-PCR检测呈阳性且免疫阳性。10天后用YHV-1攻毒,注射未感染昆虫细胞匀浆的虾在攻毒后7天内死亡率为100%,而YHV-1匀浆组直到攻毒后第9天才出现100%的死亡率。Kaplan-Meier对数秩生存分析显示两组的生存曲线有显著差异(P<0.001)。死亡率延迟的原因可能值得进一步研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d87/9852278/1c104f15d445/ga1.jpg

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