Shah Abid Ullah, Wang Zhisheng, Zheng Yating, Guo Rongli, Chen Saisai, Xu Mengwei, Zhang Chuanjian, Liu Yamei, Wang Jichun
National Research Center of Engineering and Technology for Veterinary Biologicals/Institute of Veterinary Immunology and Engineering, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
Jiangsu Co-Innovation Center for Prevention and Control of Important Animal, Infectious Diseases and Zoonoses, Yangzhou 225009, China.
Vaccines (Basel). 2022 Aug 25;10(9):1391. doi: 10.3390/vaccines10091391.
The increased virulence of infectious bursal disease virus (IBDV) is a threat to the chicken industry. The construction of novel herpesvirus of turkey-vectored (HVT) vaccines expressing VP2 of virulent IBDV may be a promising vaccine candidate for controlling this serious disease in chickens. We generated a novel infectious clone of HVT Fc-126 by inserting mini-F sequences in lieu of the glycoprotein C (gC) gene. Based on this bacterial artificial chromosome (BAC), a VP2 expression cassette containing the pMCMV IE promoter and a VP2 sequence from the virulent IBDV NJ09 strain was inserted into the noncoding area between the UL55 and UL56 genes to generate the HVT vector VP2 recombinant, named HVT-VP2-09. The recovered vectored mutant HVT-VP2-09 exhibited higher titers ( = 0.0202 at 36 h) or similar growth kinetics to the parental virus HVT Fc-126 ( = 0.1181 at 48 h and = 0.1296 at 64 h). The high reactivation ability and strong expression of VP2 by HVT-VP2-09 in chicken embryo fibroblasts (CEFs) were confirmed by indirect immunofluorescence (IFA) and Western blotting. The AGP antibodies against IBDV were detected beginning at 3 weeks post-inoculation (P.I.) of HVT-VP2-09 in 1-day-old SPF chickens. Seven of ten chickens immunized with HVT-VP2-09 were protected post-challenge (P.C.) with the virulent IBDV NJ09 strain. In contrast, all chickens in the challenge control group showed typical IBD lesions in bursals, and eight of ten died P.C. In this study, we demonstrated that (i) a novel HVT BAC with the whole genome of the Fc-126 strain was obtained with the insertion of mini-F sequences in lieu of the gC gene; (ii) HVT-VP2-09 harboring the VP2 expression cassette from virulent IBDV exhibited in vitro growth properties similar to those of the parental HVT virus in CEF cells; and (iii) HVT-VP2-09 can provide efficient protection against the IBDV NJ09 strain.
传染性法氏囊病病毒(IBDV)毒力的增强对养鸡业构成了威胁。构建表达强毒株IBDV VP2的新型火鸡疱疹病毒(HVT)载体疫苗可能是控制鸡群中这种严重疾病的一种有前景的候选疫苗。我们通过插入mini-F序列替代糖蛋白C(gC)基因,构建了一种新型的HVT Fc-126感染性克隆。基于这个细菌人工染色体(BAC),将一个包含pMCMV IE启动子和来自强毒株IBDV NJ09株VP2序列的表达盒插入到UL55和UL56基因之间的非编码区,以产生HVT载体VP2重组体,命名为HVT-VP2-09。回收的载体突变体HVT-VP2-09在36小时时表现出更高的滴度( = 0.0202),或与亲本病毒HVT Fc-126具有相似的生长动力学(48小时时 = 0.1181,64小时时 = 0.1296)。通过间接免疫荧光(IFA)和蛋白质印迹法证实了HVT-VP2-09在鸡胚成纤维细胞(CEF)中具有高再激活能力和VP2的强表达。在1日龄SPF鸡中接种HVT-VP2-09后3周开始检测到针对IBDV的AGP抗体。用HVT-VP2-09免疫的十只鸡中有七只在受到强毒株IBDV NJ09株攻击后得到保护。相比之下,攻击对照组的所有鸡在法氏囊中均出现典型的IBD病变,十只中有八只在攻击后死亡。在本研究中,我们证明:(i)通过插入mini-F序列替代gC基因,获得了具有Fc-126株全基因组的新型HVT BAC;(ii)携带来自强毒株IBDV的VP2表达盒的HVT-VP2-09在CEF细胞中的体外生长特性与亲本HVT病毒相似;(iii)HVT-VP2-09可以提供针对IBDV NJ09株的有效保护。