重组病毒活载体疫苗是一种能够有效激活特异性和非特异性免疫、可多联多价、安全性好的新型疫苗。动物α疱疹病毒拥有较大的基因组，含有多个不影响病毒复制的非必需区，能够插入接受外源基因并表达相应抗原蛋白；同时具有较广泛的宿主谱，能够在宿主体内复制并持续刺激动物产生对抗相应病原的免疫力，是作为重组病毒活载体疫苗的理想载体。随着基因编辑技术的发展，可通过多种方法构建能够表达外源基因的重组病毒。目前以动物α疱疹病毒为载体的重组病毒活载体疫苗研究已经涉及禽类、猪、牛、羊、伴侣动物等，目前成功构建的多株重组动物α疱疹病毒能免疫后可使动物同时获得对多种疾病的免疫。本文总结了重组动物α疱疹病毒构建方法、外源基因的引入和表达以及动物α疱疹病毒活载体疫苗免疫作用三个方面的内容，包括了最新的基因编辑技术、不同的构建策略及其优缺点、外源基因的选择、插入形式和位点等，并介绍了各种动物α疱疹病毒活载体疫苗的最新研究进展，旨在为新型动物α疱疹病毒活载体疫苗的研究和开发提供一定的参考。

Many mammalian herpes viruses utilize heparan sulfate (HS) moieties present on cell surface proteoglycans as receptors for cell entry, and this process also requires viral glycoprotein C (gC) homologues.  However, our understanding of the role of gC in facilitating attachment of other alpha-herpes viruses such as the duck plague virus (DPV) remains preliminary.  To study the role of gC during DPV infection, we used a gC-deleted mutant virus (DPV-ΔgC-EGFP).  Examination of the viral copy number by real-time PCR, as well as time course studies of viral adsorption and proliferation revealed that gC was involved in the viral binding to the cell surface.  The affinity of viral glycoproteins (gB-DPV, gC-DPV, and gE-DPV) to HS was assessed using a prokaryotic expression system and HiTrap^TM Heparin HP column chromatography.  In addition, to confirm that gC played a role in the interaction between DPV and HS, viruses were treated with the HS analogue heparin and host cells were treated with its inhibitors heparinase prior to exposure to DPV-ΔgC-EGFP or wild-type strain Chinese virulent duck plague virus (DPV-CHv).  The effects of heparin and heparinase on virus infectivity demonstrated that function of gC on viral adsorption is independent of interactions between gC and heparin sulfate on cell surface.  All in all, this study demonstrated that the gC of DPV can mediate viral adsorption in an HS-independent manner, which distinguish it from the gC of some other alpha-herpes viruses.  Future studies will be required to identify the receptors involved in gC protein binding to cells.  This work provides us a foundation for further studies of examining the roles of gC in the adsorption during duck plague virus infection.

    CD8, a glycoprotein on the surface of T cells, is involved in the defense against viral infection and plays significant roles in antigen presentation and in the antiviral immune response. CD8 is composed of two chains. Of these, the CD8α chain was chosen for the detection because it involved in both the CD8αα homodimer and the CD8αβ heterodimer. Here, we established a double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for specific detection of goose CD8α (goCD8α). The results showed that the optimal coated antibody and antigen dilutions were 1:50 (the antibody titer was 1:12 800) and 1:32 (0.3 ng mL^–1), respectively, while the optimal capture antibody and horseradish peroxidase (HRP)-labelled goat anti-rabbit IgG dilutions were 1:50 (the antibody titer was 1:51 200) and 1:4 000 (the antibody titer was 1:5 000), respectively. The optimal blocking buffer was 5% bovine serum albumin (BSA). The best incubating condition was overnight at 4°C, the best blocking time was 120 min and the best anti-capture antibody working time was 150 min. In addition, the minimum dose detectable by DAS-ELISA was 5×10–3 ng mL^–1. Most importantly, goCD8α expression levels in goose spleen mononuclear cells (MNCs) post-Goose parvoviruse (GPV) infection were found to be significantly up-regulated using the DAS-ELISA method, which was consistent with previous results obtained using real-time quantitative PCR. In conclusion, the DAS-ELISA method reported here is a novel, specific technique for the clinical detection of goCD8α.