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In this work, we have developed spores that could be used to vaccinate against<jats:italic>Clostridium perfringens<\/jats:italic>alpha toxin and that could be used to protect against gas gangrene in humans and necrotic enteritis in poultry. The primary active agent in both cases is alpha toxin. A carboxy-terminal segment of the alpha toxin gene (<jats:italic>cpa<\/jats:italic>) fused to the glutathione-<jats:italic>S<\/jats:italic>-transferase (GST) gene was cloned in<jats:italic>B. subtilis<\/jats:italic>such that the encoded GST-Cpa<jats:sub>247-370<\/jats:sub>polypeptide had been expressed in the following three different ways: expression in the vegetative cell, expression on the surface of the spore coat (fused to the CotB spore coat protein), and a combined approach of spore coat expression coupled with expression in the vegetative cell. Mice immunized orally or nasally with three doses of recombinant spores that carried GST-Cpa<jats:sub>247-370<\/jats:sub>on the spore surface showed the most striking responses. This included seroconversion with anti-Cpa<jats:sub>247-370<\/jats:sub>-specific immunoglobulin G (IgG) responses in their sera, a Th2 bias, and secretory IgA responses in saliva, feces, and lung samples. Neutralizing IgG antibodies to alpha toxin were detected using in vitro and in vivo assays, and a toxin challenge established protection. Mice immunized nasally or orally with recombinant spores were protected against a challenge with 12 median lethal doses of alpha toxin. Existing use of spores as competitive exclusion agents in animal feeds supports their use as a potentially economical and heat-stable vaccine for the poultry industry.<\/jats:p>","DOI":"10.1128\/iai.00686-08","type":"journal-article","created":{"date-parts":[[2008,9,9]],"date-time":"2008-09-09T00:33:56Z","timestamp":1220920436000},"page":"5257-5265","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":88,"title":["Recombinant<i>Bacillus subtilis<\/i>Expressing the<i>Clostridium perfringens<\/i>Alpha Toxoid Is a Candidate Orally Delivered Vaccine against Necrotic Enteritis"],"prefix":"10.1128","volume":"76","author":[{"given":"Tran H.","family":"Hoang","sequence":"first","affiliation":[{"name":"School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey TW20 0EX, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huynh A.","family":"Hong","sequence":"additional","affiliation":[{"name":"School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey TW20 0EX, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Graeme C.","family":"Clark","sequence":"additional","affiliation":[{"name":"Defence Science and Technology Laboratory, Porton Down, Salisbury, Wiltshire SP4 0JQ, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard W.","family":"Titball","sequence":"additional","affiliation":[{"name":"School of Biosciences, University of Exeter, Exeter, Devon EX4 4QD, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Simon M.","family":"Cutting","sequence":"additional","affiliation":[{"name":"School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey TW20 0EX, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.1995.tb02234.x"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.2307\/1591716"},{"key":"e_1_3_2_4_2","doi-asserted-by":"crossref","first-page":"3448","DOI":"10.4049\/jimmunol.138.10.3448","volume":"138","year":"1987","unstructured":"Balloul, J.-M., J.-M. 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