Chambers M A, Stagg D, Gavier-Widén D, Lowrie D, Newell D, Hewinson R G
Department of Bacterial Diseases, Veterinary Laboratories Agency Weybridge, Addlestone, Surrey, UK.
Res Vet Sci. 2001 Oct;71(2):119-26. doi: 10.1053/rvsc.2001.0498.
Nucleic acid (DNA) vaccination against tuberculosis in the European badger (Meles meles) is one approach to addressing the escalating problem of bovine tuberculosis in Great Britain. The aim of vaccination is to reduce the burden of tuberculosis within the badger population and the shedding of Mycobacterium bovis to levels that would break the transmission of infection to cattle. To this end, the vaccine would be required to limit the amount of disseminated tuberculosis in the badger, especially dissemination to the kidney from where M. bovis can be shed in the urine. A promising candidate DNA vaccine encoding a 26 kDa major antigen (MPB83) of M. bovis was evaluated in a mouse model of disseminated M. bovis infection. Using the DNA vaccine, protection against infection of the kidney was found to be greater than that achieved with the current live vaccine, Bacille Calmette-Guerin (BCG). Kidney tissue and skeletal muscle from the badger was used to derive primary cell cultures in which to examine the expression of MPB83 following transfection with the DNA vaccine. Kidney cortex gave rise to a monotypic culture of epithelial cells whilst the muscle gave rise to a mixed culture of fibroblasts and myoblasts. During culture the myoblasts differentiated into multinucleated myotubes, verified by immunofluorescent detection of mammalian desmin. Successful expression of MPB83 by transfected epithelial and myotube cells was confirmed by immunofluorescence using a monoclonal antibody specific to the protein. These observations fulfil the early requirements for the development of a DNA vaccine for badger tuberculosis.
针对欧洲獾(鼬獾)进行核酸(DNA)疫苗接种是解决英国牛结核病问题不断升级的一种方法。疫苗接种的目的是减轻獾群体内的结核病负担,并将牛分枝杆菌的排菌量降低到能够阻断向牛传播感染的水平。为此,需要疫苗限制獾体内播散性结核病的数量,尤其是限制向肾脏的播散,因为牛分枝杆菌可从肾脏随尿液排出。在牛分枝杆菌播散性感染的小鼠模型中评估了一种有前景的候选DNA疫苗,该疫苗编码牛分枝杆菌的一种26 kDa主要抗原(MPB83)。使用该DNA疫苗,发现对肾脏感染的保护作用大于目前使用的活疫苗卡介苗(BCG)。利用獾的肾脏组织和骨骼肌建立原代细胞培养物,用于检测用DNA疫苗转染后MPB83的表达。肾皮质产生了上皮细胞的单一型培养物,而肌肉产生了成纤维细胞和平滑肌细胞的混合培养物。在培养过程中,平滑肌细胞分化为多核肌管,通过免疫荧光检测哺乳动物结蛋白得以证实。使用针对该蛋白的单克隆抗体通过免疫荧光法证实了转染的上皮细胞和肌管细胞成功表达了MPB83。这些观察结果满足了开发用于獾结核病DNA疫苗的早期要求。