Cloning and characterization of Treponema hyodysenteriae antigens and protection in a CF-1 mouse model by immunization with a cloned endoflagellar antigen

Abstract

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 endoflageliar 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. 3808 on August 5, 2020 by guest http://iai.asm.org/ Downloaded from

Production and characterization of monoclonal antibodies against Serpulina hyodysenteriae and S. innocens and their use in serotyping

“…The diagnosis of swine dysentery is based on herd history, clinical signs, histopathological examination, and isolation of S. hyodysenteriae from feces (13). To date, nine serotypes and nine serogroups have been proposed, and serotypes 1, 2, 8, and 9 are most prevalent in North America (6,10,21). Although S. hyodysenteriae is the most common causative agent of swine dysentery (12,13), another species of spirochetes called Serpulina innocens, isolated from apparently healthy swine and those with postweaning scours, may also be involved in the pathogenesis of swine dysentery (16,27,28).…”

“…S. hyodysenteriae is known to be antigenically heterogenous, with different serotypes showing different geographical distributions. To date, nine serotypes and nine serogroups have been proposed (6,10,13,21). The most frequently encountered strains isolated from clinical cases of swine dysentery belong to serotypes 1 and 2 in the United States, serotypes 8 and 9 in Quebec, serotypes 4 and 7 in Europe, and serogroups A, B, and E in South Australia (6,19,21).…”

Cloning and DNA sequence analysis of a Serpulina (Treponema) hyodysenteriae gene encoding a periplasmic flagellar sheath protein

“…These proteins are probably components of the flagella (17). Furthermore, Boyden et al (2) showed that a cloned flagellar protein of S. hyodysentenae was protective in a mouse model of swine dysentery.…”