Arts Joop A J, Taverne-Thiele Anja J, Savelkoul Huub F J, Rombout Jan H W M
Cell Biology and Immunology Group, Wageningen University, Wageningen, The Netherlands.
Fish Shellfish Immunol. 2007 Jul;23(1):164-70. doi: 10.1016/j.fsi.2006.10.003. Epub 2006 Oct 24.
White spot syndrome virus (WSSV) has been a major cause of shrimp mortality in aquaculture worldwide in the past decades. In this study, WSSV infection (by immersion) and behaviour recruitment of haemocytes is investigated in gills and midgut, using an antiserum against the viral protein VP28 and a monoclonal antibody recognising haemocytes (WSH8) in a double immunohistochemical staining and in addition transmission electron microscopy was applied. More WSH 8(+) haemocytes were detected at 48 and 72 h post-infection in the gills of infected shrimp compared to uninfected animals. Haemocytes in the gills and midgut were not associated with VP28-immunoreactivity. In the gills many other cells showed virus replication in their nuclei, while infected nuclei in the gut cells were rare. Nevertheless, the epithelial cells in the midgut showed a clear uptake of VP28 and accumulation in supranuclear vacuoles (SNV) at 8h post-infection. However, epithelial nuclei were never VP28-immunoreactive and electron microscopy study suggests degradation of viral-like particles in the SNV. In contrast to the gills, the midgut connective tissue shows a clear increase in degranulation of haemocytes, resulting in the appearance of WSH8-immunoreactive thread-like material at 48 and 72 h post-infection. These results indicate recruitment of haemocytes upon immersion infection in the gills and degranulation of haemocytes in less infected organs, like the midgut.
在过去几十年里,白斑综合征病毒(WSSV)一直是全球水产养殖中虾类死亡的主要原因。在本研究中,使用抗病毒蛋白VP28的抗血清和识别血细胞的单克隆抗体(WSH8),通过双重免疫组织化学染色,对鳃和中肠中WSSV感染(通过浸泡)和血细胞的行为募集进行了研究,此外还应用了透射电子显微镜。与未感染的动物相比,在感染虾的鳃中,感染后48小时和72小时检测到更多的WSH 8(+)血细胞。鳃和中肠中的血细胞与VP28免疫反应性无关。在鳃中,许多其他细胞在其细胞核中显示病毒复制,而肠道细胞中的感染细胞核很少见。然而,中肠上皮细胞在感染后8小时显示出对VP28的明显摄取并在核上空泡(SNV)中积累。然而,上皮细胞核从未有VP28免疫反应性,电子显微镜研究表明SNV中病毒样颗粒降解。与鳃相反,中肠结缔组织显示血细胞脱颗粒明显增加,导致在感染后48小时和72小时出现WSH8免疫反应性丝状物质。这些结果表明,浸泡感染后鳃中血细胞被募集,而在感染较轻的器官(如中肠)中血细胞发生脱颗粒。