Oral Tolerance Induction in Hemophilia B Dogs Fed with Transplastomic Lettuce.
Oral Tolerance Induction in Hemophilia B Dogs Fed with Transplastomic Lettuce.
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DOI：
10.1016/j.ymthe.2016.11.009
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发表时间：
2017-02-01
期刊：
影响因子：
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通讯作者：
Daniell H
中科院分区：
文献类型：
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作者：
Herzog RW;Nichols TC;Su J;Zhang B;Sherman A;Merricks EP;Raymer R;Perrin GQ;Häger M;Wiinberg B;Daniell H
血友病患者中的抗药物抗体使治疗变得非常复杂。通过免疫耐受诱导(ITI)方案消除它们会带来巨大的成本，而ITI对因子IX(FIX)抑制剂往往无效。此外，没有预防性的ITI方案来防止抗药抗体(ADA)的形成。使用普遍的免疫抑制是有问题的。为了解决这一紧急的未得到满足的医疗需求，我们将生物封装在植物细胞中的抗原交付给血友病B犬。在水培设施中进行了在生菜叶绿体中表达的CTB-FIX融合的商业规模生产。冻干细胞经常温保存30个月后，CTB-FIX(∼1 mg/g)稳定，具有良好的折叠、二硫键和五聚体组装。用表达CTB-FIX的冻干生菜细胞喂养4只血友病B犬中的3只，观察到免疫球蛋白G(Ig G)/抑制物和IgE的形成对静脉注射FIX的抑制作用。饲喂CTB-FIX冻干植物细胞300天后，未检测到副作用。与形成高滴度抗体进行修复的对照组相比，口服耐受治疗的狗通过静脉注射FIX显著缩短了凝血时间。商业规模的生产、稳定性、冻干细胞的长期储存以及在大型非啮齿动物疾病模型中诱导耐受性的有效性为口服耐受性和低成本生产和传递生物药物提供了新的概念。抗药物抗体使血友病的治疗大大复杂化。在这篇文章中，作者报道了一种基于口服表达生菜叶绿体中的因子IX的抗原特异性免疫耐受诱导方案。该方案的有效性和安全性被成功地抑制了血友病B犬在长期服用后的抑制性抗体形成。
Anti-drug antibodies in hemophilia patients substantially complicate treatment. Their elimination through immune tolerance induction (ITI) protocols poses enormous costs, and ITI is often ineffective for factor IX (FIX) inhibitors. Moreover, there is no prophylactic ITI protocol to prevent anti-drug antibody (ADA) formation. Using general immune suppression is problematic. To address this urgent unmet medical need, we delivered antigen bioencapsulated in plant cells to hemophilia B dogs. Commercial-scale production of CTB-FIX fusion expressed in lettuce chloroplasts was done in a hydroponic facility. CTB-FIX (∼1 mg/g) in lyophilized cells was stable with proper folding, disulfide bonds, and pentamer assembly after 30-month storage at ambient temperature. Robust suppression of immunoglobulin G (IgG)/inhibitor and IgE formation against intravenous FIX was observed in three of four hemophilia B dogs fed with lyophilized lettuce cells expressing CTB-FIX. No side effects were detected after feeding CTB-FIX-lyophilized plant cells for >300 days. Coagulation times were markedly shortened by intravenous FIX in orally tolerized treated dogs, in contrast to control dogs that formed high-titer antibodies to FIX. Commercial-scale production, stability, prolonged storage of lyophilized cells, and efficacy in tolerance induction in a large, non-rodent model of human disease offer a novel concept for oral tolerance and low-cost production and delivery of biopharmaceuticals. Anti-drug antibodies substantially complicate hemophilia treatment. In this article, authors report an antigen-specific immune tolerance induction protocol based on oral delivery of factor IX expressed in lettuce chloroplasts. Effectiveness and safety of the protocol are demonstrated by successful suppression of inhibitory antibody formation in hemophilia B dogs upon long-term delivery.