Laboratory of Emerging Infectious Diseases, Department of Frontier Veterinary Science, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima, 890-0065, Japan,
Cytotechnology. 2008 Nov;58(3):127-33. doi: 10.1007/s10616-009-9181-4. Epub 2009 Feb 12.
Lactoperoxidase (LPO) is a 78 kDa heme-containing oxidation-reduction enzyme present in milk, found in physiological fluids of mammals. LPO has an antimicrobial activity, and presumably contribute to the protective functions of milk against infectious diseases. In this study, recombinant vaccinia virus expressing bovine LPO (vv/bLPO) was constructed. In rabbit kidney (RK13) cells infected with vv/bLPO, recombinant bLPO was detected in both cell extracts and culture supernatants. Tunicamycin treatment decreased the molecular weight of recombinant bLPO, indicating that recombinant bLPO contains a N-linked glycosylation site. The replication of recombinant vaccinia viruses expressing bovine lactoferrin (vv/bLF) at a multiplicity of infection (moi) of 5 plaque-forming units (PFU)/cell was inhibited by antiviral activity of recombinant bLF, suggesting that vv/bLF has an antiviral effect against vaccinia virus. On the other hand, the replication of vv/bLPO at a moi of 5 PFU/cell was not inhibited by antiviral activity of recombinant bLPO, indicating that this recombinant virus could be used as a suitable viral vector. These results indicate that a combination of bLPO and vaccinia virus vector may be useful for medical and veterinary applications in vivo.
乳过氧化物酶（LPO）是一种 78 kDa 的含血红素氧化还原酶，存在于牛奶中，也存在于哺乳动物的生理液中。LPO 具有抗菌活性，可能有助于牛奶发挥其对传染病的保护作用。本研究构建了表达牛 LPO 的重组痘苗病毒（vv/bLPO）。在感染 vv/bLPO 的兔肾（RK13）细胞中，在细胞提取物和培养上清液中均检测到重组 bLPO。衣霉素处理降低了重组 bLPO 的分子量，表明重组 bLPO 含有一个 N-连接的糖基化位点。当感染复数（moi）为 5 个噬菌斑形成单位（PFU）/细胞时，表达牛乳铁蛋白的重组痘苗病毒（vv/bLF）的复制被重组 bLF 的抗病毒活性所抑制，表明 vv/bLF 对痘苗病毒具有抗病毒作用。另一方面，moi 为 5 PFU/cell 时，重组 bLPO 的抗病毒活性并未抑制 vv/bLPO 的复制，表明该重组病毒可用作合适的病毒载体。这些结果表明，bLPO 和痘苗病毒载体的结合可能对体内的医学和兽医应用有用。
