In recent years, Bovine herpesvirus 4 (BoHV-4) has emerged as an attractive gene delivery viral vector, mainly for vaccination purposes in the veterinary field. In the present study, a new infectious clone of the BoHV-4 genome carrying a bacterial artificial chromosome vector (BoHV-4-BAC) was developed by homologous recombination in mammalian cell culture and bacterial systems, and exploited to express a truncated form of glycoprotein D (tgD) of Bovine herpesvirus 1 (BoHV-1) (BoHV-4-tgD∆TK) as a vaccine candidate. This construct’s immunogenicity was compared to a DNA vector expressing the same antigen (pC-tgD) in a BALB/c mouse model. After the mice were immunized, total and specific antibody responses, cytokine responses, total splenocyte cells proliferation/cytotoxicity, and virus neutralization assays were conducted to analyze the immune response elicited by both constructs. Mice from both vaccine groups developed significant humoral and cellular immune responses after a booster dose regime was conducted on day 28 post-injection. In almost all immunological assays, BoHV-4-tgDΔTK induced as high an immune response as pC-tgD. In both vaccine constructs, neutralizing antibodies were a significant determining factor in protection against BoHV-1, even after the first injection. We conclude that a BoHV-4-based viral vector offers an effective immunization strategy as an alternative to DNA-based immunization platforms, at least to combat BoHV-1.

近年来，牛疱疹病毒 4 (BoHV-4) 已成为一种有吸引力的基因传递病毒载体，主要用于兽医领域的疫苗接种目的。在本研究中，携带细菌人工染色体载体 (BoHV-4-BAC) 的 BoHV-4 基因组的新感染性克隆是通过哺乳动物细胞培养和细菌系统中的同源重组开发的，并用于表达截短形式的糖蛋白D (tgD) 牛疱疹病毒 1 (BoHV-1) (BoHV-4-tgD∆TK) 作为候选疫苗。将该构建体的免疫原性与在 BALB/c 小鼠模型中表达相同抗原 (pC-tgD) 的 DNA 载体进行比较。小鼠免疫后，总和特异性抗体反应、细胞因子反应、总脾细胞增殖/细胞毒性、进行病毒中和试验以分析由两种构建体引起的免疫反应。在注射后第 28 天进行加强剂量方案后，来自两个疫苗组的小鼠都产生了显着的体液和细胞免疫反应。在几乎所有免疫学检测中，BoHV-4-tgDΔTK 诱导的免疫反应与 pC-tgD 一样高。在两种疫苗构建体中，中和抗体是预防 BoHV-1 的重要决定因素，即使在第一次注射后也是如此。我们得出的结论是，基于 BoHV-4 的病毒载体提供了一种有效的免疫策略，可作为基于 DNA 的免疫平台的替代方案，至少可以对抗 BoHV-1。在注射后第 28 天进行加强剂量方案后，来自两个疫苗组的小鼠都产生了显着的体液和细胞免疫反应。在几乎所有免疫学检测中，BoHV-4-tgDΔTK 诱导的免疫反应与 pC-tgD 一样高。在两种疫苗构建体中，中和抗体是预防 BoHV-1 的重要决定因素，即使在第一次注射后也是如此。我们得出的结论是，基于 BoHV-4 的病毒载体提供了一种有效的免疫策略，可作为基于 DNA 的免疫平台的替代方案，至少可以对抗 BoHV-1。在注射后第 28 天进行加强剂量方案后，来自两个疫苗组的小鼠都产生了显着的体液和细胞免疫反应。在几乎所有免疫学检测中，BoHV-4-tgDΔTK 诱导的免疫反应与 pC-tgD 一样高。在两种疫苗构建体中，中和抗体是预防 BoHV-1 的重要决定因素，即使在第一次注射后也是如此。我们得出的结论是，基于 BoHV-4 的病毒载体提供了一种有效的免疫策略，可作为基于 DNA 的免疫平台的替代方案，至少可以对抗 BoHV-1。