College of Animal Science and Veterinary Medicine, Henan Agricultural University, Wenhua Road 95#, 450002 Zhengzhou, Henan Province, People's Republic of China.
Vaccine. 2010 Nov 29;28(51):8112-9. doi: 10.1016/j.vaccine.2010.09.106. Epub 2010 Oct 14.
Infectious bronchitis virus (IBV) poses a major threat to the chicken industry worldwide. In this study, we developed a recombinant fowlpox virus (rFPV) vaccine expressing the IBV S1 gene and chicken interleukin-18 gene (IL-18), rFPV-S1/IL18. Recombinant plasmid pSY-S1/IL18 was constructed by cloning chicken IL-18 into fowlpox virus transfer plasmid containing S1 gene and transfected into the chicken embryo fibroblasts cell pre-infected with S-FPV-017 to generate rFPV-S1/IL18. Expression of the recombinant proteins was confirmed by RT-PCR and IFA. We also constructed the recombinant fowlpox virus rFPV-S1 without IL-18. One-day-old chickens were vaccinated by wing-web puncture with the two rFPVs, and the induced humoral and cellular responses were evaluated. There was a significant difference in ELISA antibody levels (P<0.05) elicited by either rFPV-S1 or rFPV-S1/IL18. The ratios of CD4(+) to CD8(+) in chickens immunized with rFPV-S1/IL18 were significantly higher (P<0.05) than in those immunized with rFPV-S1. All chickens immunized with rFPV-S1/IL18 were completely protected (20/20) after challenge with the virulent IBV HN99 strain 43 days after immunization, while only 15 out of 20 of the chickens immunized with the rFPV-S1 were protected. Our results show that the protective efficacy of the rFPV-S1 vaccine could be enhanced significantly by simultaneous expression of IL-18.
传染性支气管炎病毒(IBV)对全球家禽业构成重大威胁。在本研究中,我们开发了一种表达 IBV S1 基因和鸡白细胞介素-18 基因(IL-18)的重组禽痘病毒(rFPV)疫苗,rFPV-S1/IL18。通过将鸡白细胞介素-18 克隆到含有 S1 基因的禽痘病毒转移质粒中,并转染到预先感染 S-FPV-017 的鸡胚成纤维细胞中,构建了重组质粒 pSY-S1/IL18,从而生成 rFPV-S1/IL18。通过 RT-PCR 和 IFA 确认了重组蛋白的表达。我们还构建了不含有白细胞介素-18 的重组禽痘病毒 rFPV-S1。通过翅膀划痕对 1 日龄雏鸡接种两种 rFPV,评估诱导的体液和细胞反应。rFPV-S1 或 rFPV-S1/IL18 引起的 ELISA 抗体水平有显著差异(P<0.05)。用 rFPV-S1/IL18 免疫的鸡的 CD4(+)与 CD8(+)的比值明显高于用 rFPV-S1 免疫的鸡(P<0.05)。在免疫后 43 天用强毒 IBV HN99 株攻毒后,所有用 rFPV-S1/IL18 免疫的鸡均完全保护(20/20),而用 rFPV-S1 免疫的鸡中有 15 只(20/20)得到保护。我们的结果表明,同时表达白细胞介素-18 可显著增强 rFPV-S1 疫苗的保护效力。