塞内卡病毒A完整病毒和空衣壳的分离及抗体应答分析

doi:10.11843/j.issn.0366-6964.2024.10.027

摘要/Abstract

摘要：

塞内卡病毒A (Senecavirus A, SVA)在复制过程中可形成完整病毒和空衣壳，它们的分离条件和抗体应答差异目前还是未知的。本研究利用盐酸胍形成SVA空衣壳，通过优化不同的密度梯度、介质、转速、时间条件进行超速离心，获得分离SVA完整病毒和空衣壳的最佳条件，并按SVA完整病毒12.5 μg·只^-1，空衣壳12.5 μg·只^-1肌肉注射免疫BALB/c小鼠，免疫后1~7周检测特异性抗体和中和抗体。结果显示，SVA的MOI=1时感染细胞2 h加入100 mmol·L^-1盐酸胍可以使SVA完整病毒全部形成空衣壳；在10%~50%氯化铯，36 000 r·min^-1，离心2.5 h是区分SVA完整病毒和空衣壳的最佳条件；而且SVA完整病毒与空衣壳诱导小鼠的特异性抗体和中和抗体水平无显著差异(P=ns)。本研究为塞内卡病毒A完整病毒和空衣壳的分离纯化、重组SVA病毒样颗粒疫苗的开发提供参考。

关键词: 塞内卡病毒A, 完整病毒, 空衣壳, 分离条件优化, 免疫原性

Abstract:

Senecavirus A (SVA) has the ability to form intact virions and empty capsids during replication; however, the conditions for isolating them and their differences in antibody response are still unknown. In this study, we utilized guanidine hydrochloride to generate SVA empty capsids and determined the optimal conditions for separating SVA intact virions and empty capsids. This was achieved through ultracentrifugation using different density gradients, media, rotation rates, and durations. Subsequently, SVA intact virions and empty capsids were intramuscularly immunized into BALB/c mice at a dosage of 12.5 μg per mouse. Specific and neutralizing antibodies were then monitored 1-7 weeks post-immunization. The findings indicated that the addition of 100 mmol·L^-1 guanidine HCl for 2 hours in infected cells at MOI=1 of SVA resulted in the complete formation of empty capsids. The most effective method for distinguishing SVA intact virions from empty capsids was centrifugation at 36 000 r·min^-1 for 2.5 hours, using a cesium chloride gradient ranging from 10% to 50%. Furthermore, the specific and neutralizing antibody levels for SVA intact virions were comparable to those of empty capsids. This study provides a new reference for the isolation and purification of the complete and empty capsid of Seneca virus A, as well as for the development of recombinant SVA virus-like particle vaccines.

Key words: Senecavirus A, intact particle, empty capsid, separation conditions, immunogenicity

中图分类号:

S852.659.6

李梅, 穆素雨, 董虎, 李硕, 潘颂佳, 郭慧琛, 孙世琪. 塞内卡病毒A完整病毒和空衣壳的分离及抗体应答分析[J]. 畜牧兽医学报, 2024, 55(10): 4562-4570.

Mei LI, Suyu MU, Hu DONG, Shuo LI, Songjia PAN, Huichen GUO, Shiqi SUN. Isolation, Purification and Immunogenicity Evaluation of Senecavirus A Intact Particle and Empty Capsid[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(10): 4562-4570.

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