Coeurdacier Jean-Luc, Laporte Florent, Pepin Jean-François
IFREMER, Route de Maguelone, 34 250, Palavas les Flots, France.
Fish Shellfish Immunol. 2003 May;14(5):435-47. doi: 10.1006/fsim.2002.0449.
Four synthetic peptides of 15 amino acids (aa), corresponding to sequences of the nodavirus DIEV RNA(2) protein, were chosen to test their potential immunogenicity in sea bass. Two of these included the N or C terminal regions (N-ter or C-ter) and the sequences of the others contained a potential external site (aa 127-140: Lp1 and as 266-279: Lp2). Two heat inactivated strains of nodavirus (HI Sb1 and HI Sb2), were used as positive controls and the carrier (KLH) as a negative control. ELISAs were performed to quantify serum antibodies specific to nodavirus, to peptides, and to the carrier in order to monitor their immunogenicity. All the fish reacted to the peptides C-Ter, Lp1 and Lp2 but only 55% of animals injected with N-ter produced specific antibodies. The proportion of fish that produced antibodies that cross reacted with nodavirus was very different with regard to the antigen injected: HI Sb1=88%; HI Sb2=85%; N-ter=38%; C-ter=27%. Protection against nodavirus was investigated by challenging the fish with a virulent viral suspension. The results showed that heat-inactivated Nodavirus protect fish and the N-ter peptide is a potential protective peptide. This initial approach showed that although vaccinating fish with peptides is possible, the tools and strategies of the research used in this field still need to be adapted to fish.
选择了4条由15个氨基酸组成的合成肽,其对应于诺达病毒DIEV RNA(2)蛋白的序列,以测试它们在海鲈中的潜在免疫原性。其中两条包含N端或C端区域(N端或C端),另外两条的序列包含一个潜在的外部位点(氨基酸127 - 140:Lp1和氨基酸266 - 279:Lp2)。两种热灭活的诺达病毒株(HI Sb1和HI Sb2)用作阳性对照,载体(KLH)用作阴性对照。进行酶联免疫吸附测定(ELISA)以定量针对诺达病毒、肽和载体的血清特异性抗体,以监测它们的免疫原性。所有鱼对肽C端、Lp1和Lp2均有反应,但仅55%注射N端的动物产生了特异性抗体。产生与诺达病毒交叉反应抗体的鱼的比例因注射的抗原而异:HI Sb1 = 88%;HI Sb2 = 85%;N端 = 38%;C端 = 27%。通过用强毒病毒悬液攻击鱼来研究对诺达病毒的保护作用。结果表明,热灭活的诺达病毒可保护鱼,N端肽是一种潜在的保护肽。这一初步方法表明,虽然用肽给鱼接种疫苗是可行的,但该领域所使用的研究工具和策略仍需针对鱼进行调整。