Lin Huixing, Ma Zhe, Chen Lei, Fan Hongjie
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China.
Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, China.
Front Vet Sci. 2017 May 30;4:81. doi: 10.3389/fvets.2017.00081. eCollection 2017.
Classical swine fever (CSF) is a highly contagious and serious viral disease that affects the pig industry worldwide. The glycoprotein E2 of the classical swine fever virus (CSFV) can induce neutralizing antibodies, and it is widely used for novel vaccine development. To explore the development of a vaccine against CSFV infections, the gene of glycoprotein E2 was inserted into the swinepox virus (SPV) genome by homologous recombination. The culture titers of rSPV-E2 remained at about 4.3 × 10 TCID for more than 60 passages in PK15 and swine testis cell lines. The rSPV-E2 could not be replicated in Vero, MDBK or other non-porcine cell lines. After two to three passages, the SPV specific gene of rSPV-E2 could not been detected in the non-porcine cell culture. To evaluate the immunogenicity of rSPV-E2, 20 CSFV seronegative minipigs were immunized with rSPV-E2, a commercial C-strain vaccine, wild-type SPV (wtSPV; negative control), or PBS (a no-challenge control). After challenge with CSFV, pigs in the rSPV-E2-immunized group showed significantly shorter fever duration compared with the wtSPV-treated group ( < 0.05). E2-specific antibodies in the rSPV-E2-immunized group increased dramatically after vaccination and increased continuously over time. CSFV genomic copies in the serum of rSPV-E2-immunized pigs were significantly less compared with the wtSPV-treated group at all time points after challenge ( < 0.01). Significant reduction in gross lung lesion scores, histopathological liver, spleen, lung, and kidney lesion scores were noted in the rSPV-E2-immunized group compared with the wtSPV-treated group ( < 0.01). The results suggested that the recombinant rSPV-E2 provided pigs with significant protection from CSFV infections; thus, rSPV-E2 offers proof of principle for the development of a vaccine for the prevention of CSFV infections in pigs.
经典猪瘟(CSF)是一种高度传染性的严重病毒性疾病,影响着全球养猪业。经典猪瘟病毒(CSFV)的糖蛋白E2可诱导中和抗体,被广泛用于新型疫苗的研发。为探索针对CSFV感染的疫苗研发,通过同源重组将糖蛋白E2基因插入猪痘病毒(SPV)基因组。重组猪痘病毒rSPV-E2在PK15和猪睾丸细胞系中传代60多代后,培养滴度维持在约4.3×10 TCID。rSPV-E2不能在Vero、MDBK或其他非猪细胞系中复制。传代两到三次后,在非猪细胞培养物中检测不到rSPV-E2的SPV特异性基因。为评估rSPV-E2的免疫原性,用rSPV-E2、市售C株疫苗、野生型SPV(wtSPV;阴性对照)或PBS(无攻毒对照)对20头CSFV血清阴性小型猪进行免疫。用CSFV攻毒后,rSPV-E2免疫组猪的发热持续时间明显短于wtSPV处理组(<0.05)。rSPV-E2免疫组接种疫苗后E2特异性抗体显著增加,并随时间持续上升。攻毒后所有时间点,rSPV-E2免疫猪血清中的CSFV基因组拷贝数均显著低于wtSPV处理组(<0.01)。与wtSPV处理组相比,rSPV-E2免疫组的大体肺病变评分、组织病理学肝、脾、肺和肾病变评分显著降低(<0.01)。结果表明,重组rSPV-E2为猪提供了对CSFV感染的显著保护;因此,rSPV-E2为开发预防猪CSFV感染的疫苗提供了原理证明。