Lorenzen Ellen, Lorenzen Niels, Einer-Jensen Katja, Brudeseth Bjørn, Evensen Oystein
Danish Institute for Food and Veterinary Research, Hangøvej 2, DK-8200 Arhus N, Denmark.
Fish Shellfish Immunol. 2005 Jul;19(1):27-41. doi: 10.1016/j.fsi.2004.10.009.
The present study was performed as a time course study of fish vaccinated with 20 microg plasmid DNA vaccine encoding either the VHSV G-protein or the VHSV N-protein. Samples of the injection site were collected sequentially over a 7-week period. The study revealed an intense positive staining by immunohistochemistry for the viral G-protein mainly in the membrane of intact myocytes, most prominent by days 10-27, and with concomitant infiltration of inflammatory cells by days 13-38 that subsequently lead to a marked reduction in the number of myocytes expressing the G-protein. By immunofluorescence, infiltrating cells positive for MHC II, IgM, and C3 were demonstrated. By contrast, in fish vaccinated with the VHSV-N construct, fewer, diffusely positive myocytes were found, most prominent by days 13-38, these having a positive reaction for the N-protein mainly in the cytoplasm and variably in the membrane. N-protein positive myocytes did not attract infiltrating cells to the same degree. Positive reaction for the N-protein almost ceased by day 48 post-vaccination.
本研究是一项时间进程研究，对象是接种了编码弹状病毒出血性败血症病毒（VHSV）糖蛋白（G蛋白）或核蛋白（N蛋白）的20微克质粒DNA疫苗的鱼类。在7周的时间内，依次采集注射部位的样本。研究显示，通过免疫组织化学法，病毒G蛋白呈现强烈的阳性染色，主要出现在完整肌细胞的膜上，在第10至27天最为明显，并且在第13至38天伴有炎性细胞浸润，随后导致表达G蛋白的肌细胞数量显著减少。通过免疫荧光法，证实存在对MHC II、IgM和C3呈阳性的浸润细胞。相比之下，在接种VHSV - N构建体的鱼类中，发现较少的、呈弥漫性阳性的肌细胞，在第13至38天最为明显，这些肌细胞对N蛋白的阳性反应主要在细胞质中，在膜上的反应程度不一。N蛋白阳性的肌细胞对浸润细胞的吸引程度不同。接种疫苗后第48天，对N蛋白的阳性反应几乎停止。
