Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan, Shandong, China.
Dev Comp Immunol. 2012 Feb;36(2):442-9. doi: 10.1016/j.dci.2011.08.009. Epub 2011 Aug 27.
White spot disease caused by white spot syndrome virus (WSSV) leads to devastating losses in shrimp farming. The WSSV envelope protein VP28, can be used as subunit vaccines that can efficiently protect shrimp against WSSV disease. However, the function of the envelope protein VP19 was not confirmed, some researches found that VP19 could protect shrimp against WSSV, and other reports found it no any protection. To detect the functions of VP28 and VP19 and find a method to prevent this disease in red swamp crayfish Procambarus clarkii, we constructed the plasmid vectors pIevp28 and pIevp19, which contains the ie1 promoter and coding region of vp28 or vp19 of WSSV, respectively. The results of quantitative real-time PCR and western blot showed that the injected vectors could transcribe corresponding mRNAs and translate to the protein VP28 or VP19 in the crayfish. The vp28 or vp19 signal was detected on the third day post injection, and maintained its expression for 30days. The mortality of the crayfish with pIevp28 showed obvious decline compared with the controls (pIe and PBS injection). However, pIevp19 seems did not affect the mortality of the crayfish compared with the controls. Furthermore, only VP28 was found tightly bound to the host haemocytes under immunocytochemistry. The results suggest that the VP28 protein might protect shrimp from the virus through competitive inhibition. We also found that oral administration of Escherichia coli with pIevp28 could protect crayfish from white spot disease, but the E. coli with pIevp19 was not. Therefore, we think that oral administration of bacteria with pIevp28 is a potentially easy therapeutic way against white spot disease in aquaculture.
白斑综合征病毒(WSSV)引起的白斑病导致对虾养殖业遭受毁灭性损失。WSSV 包膜蛋白 VP28 可用作亚单位疫苗,能有效保护对虾免受 WSSV 疾病的侵害。然而,包膜蛋白 VP19 的功能尚未得到确认,一些研究发现 VP19 可以保护对虾免受 WSSV 的侵害,而其他报告则发现它没有任何保护作用。为了检测 VP28 和 VP19 的功能,并找到一种方法来预防罗氏沼虾 Procambarus clarkii 中的这种疾病,我们构建了质粒载体 pIevp28 和 pIevp19,它们分别包含 WSSV 的 ie1 启动子和 vp28 或 vp19 的编码区。实时定量 PCR 和 Western blot 的结果表明,注射的载体可以在对虾中转录相应的 mRNA 并翻译为蛋白 VP28 或 VP19。在注射后的第 3 天检测到 vp28 或 vp19 信号,并持续表达 30 天。与对照组(pIe 和 PBS 注射)相比,携带 pIevp28 的对虾死亡率明显下降。然而,与对照组相比,携带 pIevp19 的对虾死亡率似乎没有受到影响。此外,只有 VP28 在免疫细胞化学下被发现与宿主血细胞紧密结合。结果表明,VP28 蛋白可能通过竞争抑制来保护虾类免受病毒侵害。我们还发现,口服携带 pIevp28 的大肠杆菌可以保护罗氏沼虾免受白斑病的侵害,但携带 pIevp19 的大肠杆菌则不能。因此,我们认为口服携带 pIevp28 的细菌是水产养殖中防治白斑病的一种潜在的简便治疗方法。