Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
Anaerobe. 2024 Oct;89:102896. doi: 10.1016/j.anaerobe.2024.102896. Epub 2024 Aug 8.
Clostridioides difficile is the main cause of antibiotic-associated diarrhea in humans and is a major enteropathogen in several animal species. In newborn piglets, colonic lesions caused by C. difficile A and B toxins (TcdA and TcdB, respectively) cause diarrhea and significant production losses.
The present study aimed to develop two recombinant vaccines from immunogenic C-terminal fragments of TcdA and TcdB and evaluate the immune response in rabbits and in breeding sows. Two vaccines were produced: bivalent (rAB), consisting of recombinant fragments of TcdA and TcdB, and chimeric (rQAB), corresponding to the synthesis of the same fragments in a single protein. Groups of rabbits were inoculated with 10 or 50 μg of proteins adjuvanted with aluminum or 0.85 % sterile saline in a final volume of 1 mL/dose. Anti-TcdA and anti-TcdB IgG antibodies were detected in rabbits and sows immunized with both rAB and rQAB vaccines by ELISA. The vaccinated sows were inoculated intramuscularly with 20 μg/dose using a prime-boost approach.
Different antibody titers (p ≤ 0.05) were observed among the vaccinated groups of sows (rAB and rQAB) and control. Additionally, newborn piglets from vaccinated sows were also positive for anti-TcdA and anti-TcdB IgGs, in contrast to control piglets (p ≤ 0.05). Immunization of sows with the rQAB vaccine conferred higher anti-TcdA and anti-TcdB responses in piglets, suggesting the superiority of this compound over rAB.
The synthesized recombinant proteins were capable of inducing antibody titers against C. difficile toxins A and B in sows, and were passively transferred to piglets through colostrum.
艰难梭菌是人类抗生素相关性腹泻的主要病原体,也是多种动物的主要肠道病原体。在新生仔猪中,艰难梭菌 A 型和 B 型毒素(分别为 TcdA 和 TcdB)引起的结肠病变导致腹泻和严重的生产损失。
本研究旨在从 TcdA 和 TcdB 的免疫原性 C 端片段中开发两种重组疫苗,并评估其在兔和种猪中的免疫反应。生产了两种疫苗:双价(rAB),由 TcdA 和 TcdB 的重组片段组成,嵌合(rQAB),对应于在单个蛋白中合成相同片段。用铝佐剂或 0.85%无菌生理盐水将 10 或 50μg 蛋白作为佐剂加入到 1mL 终体积中,给兔分组接种。用 ELISA 法检测免疫了 rAB 和 rQAB 疫苗的兔和母猪的抗 TcdA 和抗 TcdB IgG 抗体。用 20μg/剂量对免疫母猪进行肌肉内接种,采用初免-加强免疫程序。
免疫母猪的不同抗体滴度(p≤0.05)(rAB 和 rQAB 疫苗)与对照组不同。此外,与对照组仔猪相比,来自免疫母猪的新生仔猪也对 TcdA 和 TcdB IgGs 呈阳性(p≤0.05)。rQAB 疫苗免疫母猪可诱导仔猪产生更高的抗 TcdA 和抗 TcdB 反应,表明该化合物优于 rAB。
合成的重组蛋白能够在母猪中诱导针对艰难梭菌毒素 A 和 B 的抗体滴度,并通过初乳被动传递给仔猪。