Verheije M H, Kroese M V, van der Linden I F A, de Boer-Luijtze E A, van Rijn P A, Pol J M A, Meulenberg J J M, Steverink P J G M
Department of Infectious Diseases and Food Chain Quality, Institute for Animal Science and Health, Lelystad, The Netherlands.
Vaccine. 2003 Jun 2;21(19-20):2556-63. doi: 10.1016/s0264-410x(03)00047-1.
Three porcine reproductive and respiratory syndrome virus (PRRSV) recombinants, generated by mutagenesis of an infectious cDNA clone of the Lelystad virus (LV) isolate, were tested for their safety and protective efficacy as potential PRRSV vaccines in pigs. Recombinant vABV688 contains two amino acid substitutions in the minor structural protein GP(2) resulting in improved growth on cell line CL2621; in recombinant vABV707 the region encoding the ectodomain of the major unglycosylated membrane protein M has been replaced by that of the murine lactate dehydrogenase-elevating arterivirus; recombinant vABV746 lacks the six C-terminal amino acids of the nucleocapsid protein N. First, we determined the safety of these recombinant viruses by monitoring the stability of the introduced mutations in 8-week-old pigs. We showed that the introduced genomic mutations were maintained throughout the viraemic period. Second, the protective efficacy of immunization with the recombinant viruses against challenge with a homologous and a heterologous PRRSV strain was determined in two pigs and compared with the efficacy of vABV437, a virus derived from the parental LV cDNA. The viraemia in pigs immunized with the recombinant viruses was reduced compared to pigs immunized with vABV437. In addition, the length of viraemia was reduced in the sentinel pigs that were introduced into the groups immunized with vABV746, vABV688, and vABV707, however, all of the sentinel pigs became infected. Pigs immunized with vABV707 and vABV437 were protected against challenge with homologous virus LV-Ter Huurne and transmission of the latter virus. None of the immunized pigs were protected against heterologous challenge with the virulent US isolate SDSU#73, but the vABV707- and vABV746-immunized pigs were protected against transmission of this virus from challenged pigs. In conclusion, the obtained viral recombinants are interesting candidates to be further explored for their use as vaccines against PRRSV.
通过对莱利斯塔德病毒(LV)分离株的感染性cDNA克隆进行诱变产生了三种猪繁殖与呼吸综合征病毒(PRRSV)重组体,并在猪身上测试了它们作为潜在PRRSV疫苗的安全性和保护效力。重组病毒vABV688在次要结构蛋白GP(2)中有两个氨基酸替换,使其在CL2621细胞系上生长得到改善;在重组病毒vABV707中,主要非糖基化膜蛋白M的胞外域编码区被鼠乳酸脱氢酶升高动脉病毒的相应区域取代;重组病毒vABV746缺少核衣壳蛋白N的六个C端氨基酸。首先,我们通过监测8周龄猪中引入突变的稳定性来确定这些重组病毒的安全性。我们发现引入的基因组突变在整个病毒血症期都得以维持。其次,在两头猪中测定了用重组病毒免疫后抵抗同源和异源PRRSV毒株攻击的保护效力,并与源自亲本LV cDNA的病毒vABV437的效力进行了比较。与用vABV437免疫的猪相比,用重组病毒免疫的猪的病毒血症有所减轻。此外,引入到用vABV746、vABV688和vABV707免疫的组中的哨兵猪的病毒血症持续时间缩短,然而,所有哨兵猪都被感染了。用vABV707和vABV437免疫的猪对同源病毒LV-Ter Huurne的攻击和后者病毒的传播具有抵抗力。没有一头免疫猪对强毒美国分离株SDSU#73的异源攻击具有抵抗力,但用vABV707和vABV746免疫的猪对该病毒从受攻击猪的传播具有抵抗力。总之,所获得的病毒重组体是作为抗PRRSV疫苗进一步探索的有意义的候选物。