Department of Infectious and Parasitic Diseases, Faculty of Veterinary Medicine, University of Liège, B-4000 Liège, Belgium.
Proc Natl Acad Sci U S A. 2013 May 21;110(21):E1933-42. doi: 10.1073/pnas.1216531110. Epub 2013 Apr 29.
Wildebeests carry asymptomatically alcelaphine herpesvirus 1 (AlHV-1), a γ-herpesvirus inducing malignant catarrhal fever (MCF) to several ruminant species (including cattle). This acute and lethal lymphoproliferative disease occurs after a prolonged asymptomatic incubation period after transmission. Our recent findings with the rabbit model indicated that AlHV-1 infection is not productive during MCF. Here, we investigated whether latency establishment could explain this apparent absence of productive infection and sought to determine its role in MCF pathogenesis. First, whole-genome cellular and viral gene expression analyses were performed in lymph nodes of MCF-developing calves. Whereas a severe disruption in cellular genes was observed, only 10% of the entire AlHV-1 genome was expressed, contrasting with the 45% observed during productive infection in vitro. In vivo, the expressed viral genes included the latency-associated nuclear antigen homolog ORF73 but none of the regions known to be essential for productive infection. Next, genomic conformation analyses revealed that AlHV-1 was essentially episomal, further suggesting that MCF might be the consequence of a latent infection rather than abortive lytic infection. This hypothesis was further supported by the high frequencies of infected CD8(+) T cells during MCF using immunodetection of ORF73 protein and single-cell RT-PCR approaches. Finally, the role of latency-associated ORF73 was addressed. A lack of ORF73 did not impair initial virus replication in vivo, but it rendered AlHV-1 unable to induce MCF and persist in vivo and conferred protection against a lethal challenge with a WT virus. Together, these findings suggest that a latent infection is essential for MCF induction.
角马无症状携带牛疱疹病毒 1(AlHV-1),一种诱导多种反刍动物(包括牛)恶性卡他热(MCF)的γ疱疹病毒。这种急性和致命的淋巴增生性疾病在传播后的长时间无症状潜伏期后发生。我们最近用兔模型的发现表明,在 MCF 期间 AlHV-1 感染不是生产性的。在这里,我们研究了潜伏期的建立是否可以解释这种明显的非生产性感染,并试图确定其在 MCF 发病机制中的作用。首先,对发生 MCF 的小牛的淋巴结进行了全基因组细胞和病毒基因表达分析。虽然观察到细胞基因严重紊乱,但只有 10%的整个 AlHV-1 基因组被表达,与体外生产性感染观察到的 45%形成对比。在体内,表达的病毒基因包括潜伏相关核抗原同源物 ORF73,但不包括已知对生产性感染至关重要的区域。接下来,基因组构象分析表明 AlHV-1 基本上是附加体,进一步表明 MCF 可能是潜伏感染而不是流产裂解感染的结果。使用 ORF73 蛋白的免疫检测和单细胞 RT-PCR 方法在 MCF 期间检测到感染的 CD8(+) T 细胞的高频率,进一步支持了这一假设。最后,研究了潜伏相关 ORF73 的作用。缺乏 ORF73 不会损害体内病毒的初始复制,但它使 AlHV-1 无法诱导 MCF 并在体内持续存在,并赋予对 WT 病毒的致命挑战的保护。总之,这些发现表明潜伏感染对于 MCF 的诱导是必不可少的。