一种重组益生菌，即表达产气荚膜梭菌α-毒素的干酪乳杆菌，作为一种口服疫苗候选物，可预防气性坏疽和坏死性肠炎。
A Recombinant Probiotic, Lactobacillus casei, Expressing the Clostridium perfringens α-toxoid, as an Orally Vaccine Candidate Against Gas Gangrene and Necrotic Enteritis.
机构信息
Department of Molecular Microbiology, Kerman Branch, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Kerman, Iran.
Department of Anaerobic Bacterial Vaccine Research and Production, Kerman Branch, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Kerman, Iran.
出版信息
Probiotics Antimicrob Proteins. 2018 Jun;10(2):251-257. doi: 10.1007/s12602-017-9276-8.
The alpha-toxin is one of the virulence factors of Clostridium perfringens for gas gangrene in humans and animals or necrotic enteritis in poultry. The C-terminal domain of this toxin ( cpa ) was synthesized and cloned into pT1NX vector to construct the pT1NX-alpha plasmid. This surface-expressing plasmid was electroporated into Lactobacillus casei ATCC 393, generating the recombinant L. casei strain expressing alpha-toxoid (LC-α strain). Expression of this modified alpha-toxoid was confirmed by SDS-PAGE, immunoblotting, and direct immunofluorescence microscopy. BALB/c mice, immunized orally by the recombinant LC-α strain, elicited mucosal and significantly humoral immune responses (p < 0.05) and developed a protection against 900 MLD/mL of the standard alpha-toxin. This study showed that this recombinant LC-α strain could be a promising vaccine candidate against gas gangrene and necrotic enteritis.
α-毒素是产气荚膜梭菌的毒力因子之一，可导致人类和动物的气性坏疽或家禽的坏死性肠炎。该毒素的 C 末端结构域（cpa）被合成并克隆到 pT1NX 载体中，构建了 pT1NX-α质粒。该表面表达质粒被电穿孔到干酪乳杆菌 ATCC 393 中，产生表达α类毒素的重组干酪乳杆菌（LC-α株）。通过 SDS-PAGE、免疫印迹和直接免疫荧光显微镜检查证实了这种修饰的α类毒素的表达。通过口服重组 LC-α 株免疫 BALB/c 小鼠，诱导体液和显著的黏膜免疫应答（p<0.05），并对 900 MLD/mL 的标准α毒素产生保护作用。本研究表明，这种重组 LC-α 株可能是一种有前途的抗气性坏疽和坏死性肠炎的疫苗候选株。
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