ASFV antigens selected from genotype I immunised pigs are immunogenic, but do not protect against genotype II challenge

Lynnette C. Goatley , Priscilla Tng , Laila Al-Adwani , Zoe Hargreaves , Stepan Levin , Teresa Lambe , Christopher L. Netherton
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Abstract

African swine fever virus (ASFV) has caused recent outbreaks of viral haemorrhagic fever in domestic pigs and wild boar in Africa, Asia, Europe, Oceania and North America. Control measures for African swine fever are limited in most countries to biosecurity at the farm gate followed by movement restrictions and selective or complete culling of pigs on affected premises. Modified live vaccines are being trialled in several countries, however development of safe and effective African swine fever subunit vaccines has been restricted by a poor understanding of the key antigens required for protection, particularly for the panzootic genotype II viruses. The cellular immune response is thought to be critical for protection against African swine fever and therefore to develop an effective subunit vaccine that stimulates an anti-ASFV T-cell response we screened lymphocytes from pigs which survived challenge with Georgia 2007/1. Using an overlapping peptide library corresponding to 168 annotated open reading frames and 24 potential minor open reading frames we identified seventeen proteins which strongly stimulated secretion of interferon gamma in an ELISpot assay. The phenotype of the T cells which were stimulated by these pools of peptides were then investigated by flow cytometry. Proteins stimulating predominantly CD8+ T cells were incorporated into bivalent replication deficient adenovirus vectors and tested as potential vaccine candidates in immunisation and challenge experiments in pigs.

选自基因型 I 免疫猪的 ASFV 抗原具有免疫原性,但不能抵御基因型 II 的挑战
非洲猪瘟病毒(ASFV)最近在非洲、亚洲、欧洲、大洋洲和北美洲的家猪和野猪中引发了病毒性出血热疫情。大多数国家对非洲猪瘟的控制措施仅限于在猪场门口采取生物安全措施,然后限制猪只流动,并有选择性地或彻底扑杀受影响场所的猪只。一些国家正在试用改良活疫苗,但由于对保护所需的关键抗原了解甚少,安全有效的非洲猪瘟亚单位疫苗的开发受到了限制,特别是泛祖基因 II 型病毒。细胞免疫反应被认为是预防非洲猪瘟的关键,因此,为了开发一种能激发抗非洲猪瘟 T 细胞反应的有效亚单位疫苗,我们筛选了经受佐治亚州 2007/1 病毒挑战后存活下来的猪的淋巴细胞。利用与 168 个注释开放阅读框和 24 个潜在次要开放阅读框相对应的重叠肽库,我们在 ELISpot 试验中鉴定出了 17 种能强烈刺激γ干扰素分泌的蛋白质。然后,我们用流式细胞术研究了受这些肽池刺激的 T 细胞的表型。将主要刺激 CD8+ T 细胞的蛋白质纳入双价复制缺陷腺病毒载体,并在猪的免疫和挑战实验中作为候选疫苗进行测试。
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