Centro de Investigaciones Biológicas, Departamento de Microbiología Molecular y Biología de las Infecciones, C/Ramiro de Maeztu 9, 28040 Madrid, Spain.
Fish Shellfish Immunol. 2010 Apr;28(4):562-70. doi: 10.1016/j.fsi.2009.12.006. Epub 2009 Dec 23.
DNA vaccines and oral DNA-based immunotherapy against infectious pancreatic necrosis virus (IPNV) have scarcely been studied in salmonid fish. Here, a vector with the capsid VP2 gene inserted was encapsulated in alginate microspheres to avoid the aggressive gastrointestinal conditions experienced following oral administration. Alginate microspheres were effective to protect the pDNA encoding VP2, which was expressed early in different organs of the vaccinated trout and that persisted for at least 60 days. The vaccine induces innate immune responses, raising the expression of IFN more than 10-fold relative to the fish vaccinated with the empty plasmid, at 7 and 15 days post-vaccination. Likewise, maximal expression of the IFN-induced antiviral Mx protein was recorded 15 days post-vaccination and neutralizing antibodies were also detected after 15 days, although their titre rose further at 21 days post-vaccination. Protection was high in the immunized fish, which showed around an 80% relative survival when challenged 15 and 30 days after vaccine delivery. Very low viral load with respect to the control group was detected in the vaccinated fish that survived 45 days after challenge. Thus, this study demonstrates the potential of the encapsulation technique for IPNV-DNA vaccine delivery and the relevance of the IPNV-VP2 gene for future plasmid constructs.
DNA 疫苗和口服 DNA 免疫疗法在鲑鱼鱼类中对传染性胰脏坏死病毒(IPNV)的研究甚少。在这里,将插入衣壳 VP2 基因的载体包封在海藻酸盐微球中,以避免口服后经历的具有侵袭性的胃肠道条件。海藻酸盐微球能够有效地保护编码 VP2 的 pDNA,该 pDNA 在接种鲑鱼的不同器官中早期表达,并至少持续 60 天。该疫苗可诱导先天免疫反应,与接种空质粒的鱼类相比,在接种后 7 和 15 天,IFN 的表达提高了 10 多倍。同样,IFN 诱导的抗病毒 Mx 蛋白的最大表达在接种后 15 天记录,并且在接种后 15 天也检测到了中和抗体,尽管其滴度在接种后 21 天进一步升高。免疫鱼的保护率很高,在疫苗接种后 15 和 30 天进行攻毒时,其相对存活率约为 80%。在接种疫苗后 45 天仍存活的鱼中,与对照组相比,病毒载量非常低。因此,这项研究证明了封装技术在 IPNV-DNA 疫苗传递中的潜力,以及 IPNV-VP2 基因对于未来质粒构建的重要性。