Division of Avian Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150001, PR China.
Vet Microbiol. 2013 Sep 27;166(1-2):59-67. doi: 10.1016/j.vetmic.2013.04.031. Epub 2013 May 20.
The present study was undertaken to evaluate the ability of DNA vaccination to protect chickens against reticuloendotheliosis virus (REV) infection and to determine whether codon optimization and the woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) could improve the immunogenicity of the DNA vaccines. The wild-type and codon-optimized gp90 genes of REV were cloned into pCAGGS vector, and designated as pCAGgp90 and pCAGoptigp90, respectively. Plasmids pCAGWgp90 and pCAGWoptigp90 containing WPRE were also constructed. To evaluate vaccine efficacy, 3-week-old specific pathogen free chickens were injected with the constructed plasmids twice at 3-week intervals and challenged with REV 3 weeks post boost. Plasmids pCAGoptigp90 and pCAGWgp90 elicited significantly higher humoral and cellular immune responses than pCAGgp90, while chickens immunized with pCAGWoptigp90 showed the highest immune responses among the groups. Chickens immunized with pCAGgp90, pCAGoptigp90, pCAGWgp90 or pCAGWoptigp90 had 53%, 67%, 73% or 87% protection, respectively, as evidenced by the absence of REV viremia, while the empty vector pCAGGS only conferred 13% protection against viremia. These results highlight the potential value of DNA vaccination in the prevention of REV infection and suggest that codon optimization and WPRE could increase the efficacy of DNA vaccines.
本研究旨在评估 DNA 疫苗对鸡传染性法氏囊病病毒(REV)感染的保护能力,并确定密码子优化和土拨鼠肝炎病毒转录后调控元件(WPRE)是否能提高 DNA 疫苗的免疫原性。将 REV 的野生型和密码子优化的 gp90 基因分别克隆到 pCAGGS 载体中,命名为 pCAGgp90 和 pCAGoptigp90。还构建了含有 WPRE 的质粒 pCAGWgp90 和 pCAGWoptigp90。为了评估疫苗的功效,将 3 周龄无特定病原体鸡每隔 3 周用构建的质粒注射两次,并在加强免疫后 3 周用 REV 攻毒。与 pCAGgp90 相比,pCAGoptigp90 和 pCAGWgp90 能诱导更高的体液和细胞免疫反应,而用 pCAGWoptigp90 免疫的鸡在各组中表现出最高的免疫反应。用 pCAGgp90、pCAGoptigp90、pCAGWgp90 或 pCAGWoptigp90 免疫的鸡,分别有 53%、67%、73%或 87%的保护率,无 REV 病毒血症,而空载体 pCAGGS 仅对病毒血症有 13%的保护率。这些结果突出了 DNA 疫苗在预防 REV 感染中的潜在价值,并表明密码子优化和 WPRE 可以提高 DNA 疫苗的功效。