Zimmermann Jana, Saalbach Isolde, Jahn Doreen, Giersberg Martin, Haehnel Sigrun, Wedel Julia, Macek Jeanette, Zoufal Karen, Glünder Gerhard, Falkenburg Dieter, Kipriyanov Sergey M
Novoplant GmbH, Am Schwabeplan 1b, 06466 Gatersleben, Germany.
BMC Biotechnol. 2009 Sep 11;9:79. doi: 10.1186/1472-6750-9-79.
Coccidiosis caused by protozoans of genus Eimeria is a chicken parasitic disease of great economical importance. Conventional disease control strategies depend on vaccination and prophylactic use of anticoccidial drugs. Alternative solution to prevent and treat coccidiosis could be provided by passive immunization using orally delivered neutralizing antibodies. We investigated the possibility to mitigate the parasitic infection by feeding poultry with antibody expressing transgenic crop seeds.
Using the phage display antibody library, we generated a panel of anti-Eimeria scFv antibody fragments with high sporozoite-neutralizing activity. These antibodies were expressed either transiently in agrobacteria-infiltrated tobacco leaves or stably in seeds of transgenic pea plants. Comparison of the scFv antibodies purified either from tobacco leaves or from the pea seeds demonstrated no difference in their antigen-binding activity and molecular form compositions. Force-feeding experiments demonstrated that oral delivery of flour prepared from the transgenic pea seeds had higher parasite neutralizing activity in vivo than the purified antibody fragments isolated from tobacco. The pea seed content was found to protect antibodies against degradation by gastrointestinal proteases (>100-fold gain in stability). Ad libitum feeding of chickens demonstrated that the transgenic seeds were well consumed and not shunned. Furthermore, feeding poultry with shred prepared from the antibody expressing pea seeds led to significant mitigation of infection caused both by high and low challenge doses of Eimeria oocysts.
The results suggest that our strategy offers a general approach to control parasitic infections in production animals using cost-effective antibody expression in crop seeds affordable for the animal health market.
由艾美耳属原生动物引起的球虫病是一种具有重大经济意义的鸡寄生虫病。传统的疾病控制策略依赖于疫苗接种和抗球虫药物的预防性使用。通过口服递送中和抗体进行被动免疫可为预防和治疗球虫病提供替代解决方案。我们研究了通过用表达抗体的转基因作物种子喂养家禽来减轻寄生虫感染的可能性。
利用噬菌体展示抗体文库,我们产生了一组具有高子孢子中和活性的抗艾美耳球虫单链抗体片段。这些抗体要么在农杆菌浸润的烟草叶片中瞬时表达,要么在转基因豌豆植物种子中稳定表达。对从烟草叶片或豌豆种子中纯化的单链抗体的比较表明,它们的抗原结合活性和分子形式组成没有差异。强制喂食实验表明,口服由转基因豌豆种子制备的面粉在体内比从烟草中分离的纯化抗体片段具有更高的寄生虫中和活性。发现豌豆种子成分可保护抗体不被胃肠道蛋白酶降解(稳定性提高超过100倍)。自由采食实验表明,鸡很乐意食用转基因种子,并不排斥。此外,用表达抗体的豌豆种子制备的碎片喂养家禽可显著减轻高剂量和低剂量艾美耳球虫卵囊引起的感染。
结果表明,我们的策略提供了一种通用方法,可利用在动物健康市场上负担得起的作物种子中进行具有成本效益的抗体表达来控制生产动物中的寄生虫感染。