针对不同形式猪繁殖与呼吸综合征病毒GP5重组蛋白的抗体中和活性比较

doi:10.11843/j.issn.0366-6964.2018.06.014

Abstract

Abstract:

GP5 protein of porcine reproductive and respiratory virus (PRRSV) is considered to be the main protein that inducing production of neutralizing antibodies. In order to investigate the ability of the protective immunity induced by the GP5 protein, the full-length GP5 and the truncated GP5 lacking of the signal peptide (aa 1-31) and transmembrane region (aa 66-125) were expressed using baculovirus and prokaryotic expression systems. Additionally, the truncated GP5 lacking of the signal peptide(aa 1-31) and transmembrane region (aa 66-125) were expressed using E. coli expression systems. Three anti-sera (ELISA assay antibody titre:800, 1 600, and 200 respectively) specific for each recombinant protein were derived from mice immunized with three recombinant proteins with Freund's Incomplete Adjuvant. PAM cells used in neutralization assay were obtained from PRRSV-negative healthy pigs by bronchoalveolar lavage after necropsy. PRRSV HeN-3 (the heterologous strain) and VR2332 (the homologous strain) strains were isolated in Marc-145 cells. The real-time quantitative PCR was used to detect PRRSV RNA levels in infected PAM cells. It was considered positive that virus blocking rate was higher than 70% which had been established in our laboratory. Results were as follows:1) The results showed that three anti-sera specific for three recombinant proteins did not have neutralizing activities against two strains of PRRSV (titers were < 1:2). 2) The anti-sera of PRRSV VR2332 had neutralizing activity against PRRSV VR2332 strain (titers were 1:8), and hadn't neutralizing activity against PRRSV HeN-3 strain but rather enhanced infection. Our current work showed the main neutralizing epitopes of GP5 were probably conformational epitopes and the neutralizing epitopes of PRRSV were not fully presented in the GP5 protein. The positive anti-sera of PRRSV could provide immune protection, but the immune protection occurs only in the same strain. It would mediate the ADE effect on heterologous strains on the contrary.

CLC Number:

FENG Yan, GUO Jia, XU Rui-qin, MA Si-xu, WEI Feng-ling, ZHANG Liu-jun, YANG Guo-yu, XIA Ping-an, ZHANG Gai-ping. Comparison of Neutralizing Antibodies Induced by PRRSV GP5 Recombinant Proteins in Different Forms of Truncation[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(6): 1222-1230.

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Comparison of Neutralizing Antibodies Induced by PRRSV GP5 Recombinant Proteins in Different Forms of Truncation

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