Programa de Pós-Graduação em Veterinária, Faculdade de Veterinária, Universidade Federal de Pelotas (UFPEL), Pelotas, RS, Brazil.
Centro de Desenvolvimento Tecnológico, Biotecnologia, Universidade Federal de Pelotas (UFPEL), Pelotas, RS, Brazil.
Anaerobe. 2024 Oct;89:102895. doi: 10.1016/j.anaerobe.2024.102895. Epub 2024 Aug 8.
Producing commercial bacterins/toxoids against Clostridium spp. is laborious and hazardous. Conversely, developing prototype vaccines using purified recombinant toxoids, though safe and effective, is both laborious and costly for application in production animals.
Considering that inactivated recombinant Escherichiacoli (bacterin) is a simple, cost-effective, and to be safe solution, we evaluated, for the first time, a pentavalent formulation of recombinant bacterins containing the alpha, beta, and epsilon toxins of Clostridiumperfringens and C and D neurotoxins of Clostridiumbotulinum in sheep.
Subcutaneously, 18 Texel sheep received two doses (200 μg of each antigen) of recombinant bacterin (n = 7) or purified recombinant antigens (n = 6) on days 0 and 28, while the control group (n = 5) did not receive an immunization. Sera samples from days 0 (before the 1st dose), 28 (before the 2nd dose), and 56, 84, and 112 were used for measuring IgG (indirect ELISA) and neutralizing antibodies (mouse serum neutralization).
Both formulations induced significant levels of IgG against all five toxins (p < 0.05) up to day 112, with peaks at days 28 and 56 post-immunization. The expected booster effect occurred only for the botulinum toxins. The neutralizing antibody titers were satisfactory against ETX (≥2 IU/ml for both formulations) and BoNT-D [5 IU/ml (bacterin) and 10 IU/ml (purified)].
While adjustments are required, the recombinant bacterin platform holds great potential for polyvalent vaccines due to its straightforward, safe, and cost-effective production, establishing it as a user-friendly technology for the veterinary immunobiological industry.
生产针对梭菌属的商业菌苗/类毒素既费力又危险。相反,使用纯化的重组类毒素开发原型疫苗,虽然安全有效,但对于生产动物的应用既费力又昂贵。
鉴于灭活重组大肠杆菌(菌苗)是一种简单、经济有效且安全的解决方案,我们首次评估了含有梭菌α、β和ε毒素以及梭菌 C 和 D 神经毒素的五价重组菌苗在绵羊中的制剂。
18 只特克赛尔羊在第 0 天和第 28 天接受两次(每种抗原 200μg)重组菌苗(n=7)或纯化重组抗原(n=6)的皮下接种,而对照组(n=5)未接受免疫接种。在第 0 天(第 1 剂前)、第 28 天(第 2 剂前)和第 56、84 和 112 天采集血清样本,用于测量 IgG(间接 ELISA)和中和抗体(小鼠血清中和)。
两种制剂均能诱导针对所有五种毒素的 IgG 水平显著升高(p<0.05),直至第 112 天,免疫后第 28 天和第 56 天达到峰值。仅在接种博特氏菌毒素后才出现预期的增强效应。两种制剂对 ETX 的中和抗体滴度均令人满意(均≥2 IU/ml),对 BoNT-D 的中和抗体滴度也令人满意[5 IU/ml(菌苗)和 10 IU/ml(纯化)]。
尽管需要进行调整,但由于其简单、安全且具有成本效益的生产,重组菌苗平台在多价疫苗方面具有巨大潜力,使其成为兽医免疫生物学行业的一种用户友好型技术。