Isolation and Characterization of a Protective Antigen-Containing Particle from Culture Supernatant Fluids of
            <i>Erysipelothrix rhusiopathiae</i>

White, R. R.; Verwey, W. F.

doi:10.1128/iai.1.4.380-386.1970

Cited by 21 publications

(22 citation statements)

References 11 publications

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“…It has been reported that the protective antigen of E. rhusiopathiae is also present in the culture supernatant of the organism and that antiserum against the culture supernatant fluids is protective (8,9,15,16). Although we found that SpaA.1 is also present in the culture supernatant, we do not yet know the relationship between the bound SpaA.…”

Section: Discussioncontrasting

confidence: 75%

“…Although we found that SpaA.1 is also present in the culture supernatant, we do not yet know the relationship between the bound SpaA. the protective antigens found in the culture supernatant reported so far (8,9,15,16).…”

Section: Discussionmentioning

confidence: 55%

“…It has been reported that a 200-kDa glycolipoprotein complex in the culture supernatant is a protective antigen of E. rhusiopathiae (15,16). This antigen was able to adsorb passively protective antibodies from rabbit antiserum produced by immunization with whole culture, suggesting that this molecule is a major protective antigen of the organism.…”

mentioning

confidence: 93%

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Immunological Characterization of a Protective Antigen of Erysipelothrix rhusiopathiae: Identification of the Region Responsible for Protective Immunity

1999

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The gene encoding a protective protein antigen of the gram-positive bacterium Erysipelothrix rhusiopathiae, an important veterinary pathogen responsible for erysipelas in swine and a variety of diseases in animals, was cloned and sequenced. The gene encodes a polypeptide of 597 amino acids plus a putative signal sequence of 29 amino acids, resulting in a mature protein with a molecular mass of 69,017 Da. Sequence analysis of the gene product revealed a C-terminal region composed of nine tandem repeats of 20 amino acids and a total sequence that is nearly identical to that of the 64-kDa cell surface protein (SpaA) of the bacterium. Because of this similarity, the protein was designated SpaA.1. In this study, we examined whether the SpaA.1 protein could induce protective antibodies and whether we could identify the region involved in protective immunity. Both the mature SpaA.1 protein and its C-terminal repeat region, but not the N-terminal segment, were expressed in Escherichia coli and purified as a histidine-tagged fusion recombinant protein. Rabbit antiserum raised against the mature SpaA.1 protein passively protected mice from lethal challenge with a virulent homologous strain, Fujisawa-SmR, suggesting that protection is mediated by humoral antibodies. To determine which domain of the SpaA.1 protein is responsible for the observed protection, mice were actively immunized with either the mature SpaA.1 protein or the C-terminal repeat region and then challenged with Fujisawa-SmR. The result showed that mice immunized with the mature SpaA.1 protein, but not the Cterminal repeat region, were protected, suggesting that the protection-eliciting epitope(s) is located within the N-terminal two-thirds of the SpaA.1 molecule. This was confirmed by passive immunization experiments in which the protective activity of rabbit antiserum, raised against mature SpaA.1 protein, was not abolished by absorption with the purified recombinant C-terminal repeat region. In addition, antibodies specific for the C-terminal repeat region were unable to protect mice from lethal challenge. These results show that the N-terminal two-thirds of the SpaA.1 molecule may constitute a good vaccine candidate against erysipelas.

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Section: Discussioncontrasting

confidence: 75%

Section: Discussionmentioning

confidence: 55%

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Immunological Characterization of a Protective Antigen of Erysipelothrix rhusiopathiae: Identification of the Region Responsible for Protective Immunity

1999

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“…From these results it was suggested that the pigs having the specific antibody to P64 could resist infection with the virulent strain (Fujisawa) of E. rhusiopathiae and that P64 is a cross-protective antigen, as P64 from strain Agata (serovar 5) could protect pigs against the infection caused by strain Fujisawa (serovar 1a). Such a cross-protective antigen of E. rhusiopathiae has been reported by many investigators (WHITE and VERWEY, 1970;SAWADA and TAKAHASHI, 1987a, b;SATO et al, 1995).…”

Section: Discussionsupporting

confidence: 56%

Protective Activity of the Purified Protein Antigen of Erysipelothrix rhusiopathiae in Pigs

Yamazaki

Sato

Sakakura

et al. 1999

Journal of Veterinary Medicine, Series B

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We purified the protein antigen (P64), which contains 66 and 64 kDa proteins, from the alkaline extract (AE) of whole cells of Erysipelothrix rhusiopathiae strain Agata (serovar 5) to determine the protective activity of the antigen against E. rhusiopathiae infection in pigs. The serum titre of antibody against P64 rapidly increased in pigs immunized with 500 and 100 mg of P64 and reached maximum values at 3 weeks after the first immunization (1 week after the second immunization). However, the serum antibody titres were not increased in pigs immunized with 20 mg of P64 and in nonimmunized pigs. In the pigs immunized with live cell vaccine (acriflavin-fast attenuated strain Koganei 65-0.15), the serum titres of antibody against P64 also increased at 1-2 weeks after immunization. In a pig challenge test performed on immunized and nonimmunized pigs, all nonimmunized pigs showed typical clinical signs of swine erysipelas (fever, erysipeloid, arthritis), while all pigs immunized with 500 and 100 mg of P64 and live cell vaccine showed no clinical signs of this disease. In Western blot analysis, sera from pigs immunized with P64 and live cell vaccine strongly reacted with the 64 kDa protein. In contrast, the serum from nonimmunized pigs did not react with any proteins. From these results, it was suggested that a specific antibody against the 64 kDa protein could be increased in pigs immunized with P64 or live cell vaccine and that this anti-P64 antibody has a strong protective effect against E. rhusiopathiae infection in pigs. U. S.

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“…The accompanying paper (7) reported on the isolation and characterization of a large protective antigen-containing particle or aggregate found in culture supernatant fluid of Erysipelothrix rhusiopathiae. The protective antigen present in these particles was representative of the major protective antigen of E. rhusiopathiae in that it was capable of absorbing all of the protective activity present in anti-whole culture rabbit serum.…”

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confidence: 99%

Solubilization and Characterization of a Protective Antigen of Erysipelothrix rhusiopathiae

White¹,

Verwey

1970

Infect Immun

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A particulate fraction of Erysipelothrix rhusiopathiae cultures has been subjected to butanol extraction and treatment with surface-active agents in an attempt to solubilize a protective antigen. The particulate fraction was partitioned into the butanol layer but was not solubilized. Only sodium dodecyl sulfate solubilized the particles. The soluble protective activity was not sedimented by centrifugation at 198,000 × g for 12 hr but was excluded by Sephadex G-200. Immunodiffusion studies of the soluble fraction demonstrated eight antigens, five of horse serum origin and three of E. rhusiopathiae origin. Ultracentrifugation indicated that spontaneous reaggregation occurred after removal of the sodium dodecyl sulfate. Analytical ultracentrifugation showed that 90% of the sodium dodecyl sulfatetreated material migrated as a single homogeneous 3.5 S component. The physical and biological characteristics of the protective activity suggest that the protective antigen is a glyco-lipoprotein.

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Isolation and Characterization of a Protective Antigen-Containing Particle from Culture Supernatant Fluids of Erysipelothrix rhusiopathiae

Cited by 21 publications

References 11 publications

Immunological Characterization of a Protective Antigen of Erysipelothrix rhusiopathiae: Identification of the Region Responsible for Protective Immunity

Immunological Characterization of a Protective Antigen of Erysipelothrix rhusiopathiae: Identification of the Region Responsible for Protective Immunity

Protective Activity of the Purified Protein Antigen of Erysipelothrix rhusiopathiae in Pigs

Solubilization and Characterization of a Protective Antigen of Erysipelothrix rhusiopathiae

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