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A Noncytolytic alpha Toxin Recombinant Protein Protects Turkeys Against Clostridium septicum Challenge

机译:非溶细胞α毒素重组蛋白可保护火鸡抵抗败血梭状芽胞杆菌的攻击。

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Clostridium septicum and its associated cytolytic alpha toxin, along with several other clostridial species, has been implicated as the causative agent of gangrenous dermatitis. A recombinant noncytolytic C. septicum alpha toxin (NCAT) peptide was developed for use as a vaccine and demonstrated to be safe at concentrations as high as 1 mg/ml. NCAT, used as a purified antigen, partially purified antigen, or in combination with native antigens, was compared to salt-fractionated a toxin combined with denatured C. septicum bacteria (native) in a vaccination trial. Three-day-old poults were placed into one of five groups and received two, 0.2-ml vaccinations 5 wk apart. Subcutaneous challenge with 3.2 X 10(7) log phase C. septicum resulted in 78% to 95% of the vaccinated birds surviving challenge compared to 48% of sham-injected controls. By ELISA analysis on NCAT-coated plates, birds receiving vaccines containing the recombinant NCAT peptide showed significantly higher blood serum antibody concentrations than did birds receiving vaccines containing native antigens or alum controls. Additionally, high levels of maternally transferred antibodies reactive to NCAT-purified antigens found in the pre-immune sera from naive 3-day-old poults suggest that the tertiary structure of the NCAT peptide has a high homology to the native protein structure. In conclusion, our study showed that the use of a vaccine comprised of a noncytolytic recombinant alpha toxin peptide antigen provided clinical protection equal to the use of vaccines formulated with inactivated native proteins at a reduced overall cost.
机译:败血梭状芽胞杆菌及其相关的溶细胞性α毒素以及其他几种梭菌属细菌已被认为是坏疽性皮炎的病原体。已开发出一种重组的非细胞溶解性败血梭菌α毒素(NCAT)肽用作疫苗,并已证明在浓度高达1 mg / ml时是安全的。在疫苗接种试验中,将用作纯化抗原,部分纯化抗原或与天然抗原结合的NCAT与盐分毒素与变性败血梭菌细菌(天然)结合进行了比较。将三天大的家禽分为五组,每组隔5周接受两次0.2 ml的疫苗接种。用3.2 X 10(7)对数败血梭状芽胞杆菌进行皮下攻击后,接种疫苗的禽鸟存活78%至95%,而假注射的对照组为48%。通过在NCAT包被的板上进行ELISA分析,与含有天然抗原或明矾对照的疫苗相比,接受含重组NCAT肽疫苗的禽类的血清抗体浓度明显更高。另外,高水平的母体转移抗体与幼稚的3天大的家禽在免疫前血清中发现的NCAT纯化抗原具有反应性,这表明NCAT肽的三级结构与天然蛋白结构具有高度同源性。总之,我们的研究表明,使用由非溶细胞重组α毒素肽抗原组成的疫苗可提供与使用灭活天然蛋白配制的疫苗相同的临床保护,并且降低了总成本。

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