Boyden D A, Albert F G, Robinson C S
Syntro Corporation, San Diego, California 92121.
Infect Immun. 1989 Dec;57(12):3808-15. doi: 10.1128/iai.57.12.3808-3815.1989.
We cloned genes that code for Treponema hyodysenteriae antigens into Escherichia coli with the purpose of identifying protective antigens for vaccine development. Three different genomic libraries were screened with various antisera reactive with T. hyodysenteriae antigens. The cloned antigens and corresponding native T. hyodysenteriae antigens were analyzed for molecular size, serum reactivity, solubility in sarcosine, and segregation during phase partitioning with the nonionic detergent Triton X-114. The results from these analyses suggested that the gene products were components of either the cytoplasmic membrane, periplasm, or endoflagella of T. hyodysenteriae. The cloned antigens were tested as vaccine candidates in a CF-1 mouse model of T. hyodysenteriae infection and immunity. Intraperitoneal injection of crude E. coli extracts containing cloned antigens did not protect mice from challenge. However, serum from mice injected with a crude extract of an E. coli clone which expressed an endoflagellar antigen killed T. hyodysenteriae in vitro. Partially purified preparations of this cloned endoflagellar antigen protected mice against oral challenge with both the homologous serotype (B204) and a heterologous serotype (B234) of T. hyodysenteriae. These results suggest that the endoflagellar proteins could be used as an effective subunit vaccine against T. hyodysenteriae.
我们将编码猪痢疾短螺旋体抗原的基因克隆到大肠杆菌中,目的是鉴定用于疫苗开发的保护性抗原。用与猪痢疾短螺旋体抗原反应的各种抗血清筛选了三个不同的基因组文库。对克隆的抗原和相应的天然猪痢疾短螺旋体抗原进行了分子大小、血清反应性、在肌氨酸中的溶解性以及用非离子去污剂Triton X-114进行相分配时的分离情况分析。这些分析结果表明,基因产物是猪痢疾短螺旋体细胞质膜、周质或内鞭毛的组成部分。在猪痢疾短螺旋体感染和免疫的CF-1小鼠模型中,对克隆的抗原作为候选疫苗进行了测试。腹腔注射含有克隆抗原的大肠杆菌粗提物并不能保护小鼠免受攻击。然而,注射了表达内鞭毛抗原的大肠杆菌克隆粗提物的小鼠血清在体外能杀死猪痢疾短螺旋体。这种克隆的内鞭毛抗原的部分纯化制剂可保护小鼠免受同源血清型(B204)和异源血清型(B234)猪痢疾短螺旋体的口服攻击。这些结果表明,内鞭毛蛋白可作为一种有效的抗猪痢疾短螺旋体亚单位疫苗。