Imada Y, Goji N, Ishikawa H, Kishima M, Sekizaki T
National Institute of Animal Health, Tsukuba, Ibaraki 305-0856, Japan.
Infect Immun. 1999 Sep;67(9):4376-82. doi: 10.1128/IAI.67.9.4376-4382.1999.
Erysipelothrix rhusiopathiae is a causal agent of swine erysipelas, which is of economic importance in the swine industry by virtue of causing acute septicemia, chronic arthritis, and endocarditis. However, little is known about the genetic properties of its protective antigens. Recently, a surface protective antigen (SpaA) gene was identified from serotype 2 in a mouse model. We cloned spaA from virulent strain Fujisawa (serotype 1a) and determined that the N-terminal 342 amino acids without C-terminal repeats of 20 amino acids have the ability to elicit protection in mice. Fusions of 342 amino acids of Fujisawa SpaA and histidine hexamer (HisSpa1.0) protected pigs against challenge with both serotype 1 and serotype 2, the most important serotypes in the swine industry. Pigs immunized with HisSpa1.0 reacted well with both HisSpa1.0 and intact SpaA by enzyme-linked immunosorbent assay and immunoblotting. Serum collected at the time of challenge from a pig immunized with HisSpa1. 0 markedly enhanced the in vitro phagocytic and killing activity of pig neutrophils against the bacteria. DNA sequences of protective regions of spaA genes from five strains of serotypes 1 and 2 were almost identical. The full DNA sequences also seemed to be conserved among strains of all 12 serotype reference strains harboring the spaA gene by restriction fragment length polymorphism analysis of PCR products. These results indicates that SpaA is a common protective antigen of serotypes 1 and 2 of E. rhusiopathiae in swine and will be a useful tool for development of new types of vaccines and diagnostic tools for effective control of the disease.
猪丹毒丝菌是猪丹毒的病原体,由于会引发急性败血症、慢性关节炎和心内膜炎,在养猪业中具有重要经济意义。然而,对其保护性抗原的遗传特性却知之甚少。最近,在小鼠模型中从2型菌株中鉴定出一种表面保护性抗原(SpaA)基因。我们从强毒株藤泽(血清型1a)中克隆了spaA,并确定其N端342个氨基酸(无C端20个氨基酸的重复序列)具有在小鼠中引发保护作用的能力。藤泽SpaA的342个氨基酸与组氨酸六聚体的融合蛋白(HisSpa1.0)可保护猪抵御血清型1和血清型2(养猪业中最重要的血清型)的攻击。用HisSpa1.0免疫的猪通过酶联免疫吸附测定和免疫印迹法与HisSpa1.0和完整的SpaA反应良好。在攻毒时从用HisSpa1.0免疫的猪采集的血清显著增强了猪中性粒细胞对细菌的体外吞噬和杀伤活性。血清型1和2的5个菌株的spaA基因保护区域的DNA序列几乎相同。通过对PCR产物的限制性片段长度多态性分析,在所有携带spaA基因的12个血清型参考菌株中,完整的DNA序列似乎也保守。这些结果表明,SpaA是猪丹毒丝菌血清型1和2的共同保护性抗原,将成为开发新型疫苗和诊断工具以有效控制该病的有用工具。