Abstract:
A short mouse-specific peptide from zona pellucida 3 (mZP3, amino acids 328-342) has been shown to be associated with antibody-mediated contraception. In this study, we investigated the production of mZP3 in the plant, as an orally applicable host, and examined the immunogenicity of this small peptide in the BALB/c mouse model. The mZP3 peptide was inserted into the major immunodominant region of the hepatitis B core antigen and was produced in Nicotiana benthamiana plants via Agrobacterium-mediated transient expression. Soluble HBcAg-mZP3 accumulated at levels up to 2.63 mg/g leaf dry weight (LDW) containing ~172 µg/mg LDW mZP3 peptide. Sucrose gradient analysis and electron microscopy indicated the assembly of the HBcAg-mZP3 virus-like particles (VLPs) in the soluble protein fraction. Subcutaneously administered mZP3 peptide displayed on HBcAg VLPs was immunogenic in BALB/c mice at a relatively low dosage (5.5 µg mZP3 per dose) and led to the generation of mZP3-specific antibodies that bound to the native zona pellucida of wild mice. Oral delivery of dried leaves expressing HBcAg-mZP3 also elicited mZP3-specific serum IgG and mucosal IgA that cross-reacted with the zona pellucida of wild mice. According to these results, it is worthwhile to investigate the efficiency of plants producing HBcAg-mZP3 VLPs as immunogenic edible baits in reducing the fertility of wild mice through inducing antibodies that cross-react to the zona pellucida.
摘要:
来自透明带3的短小鼠特异性肽(mZP3,氨基酸328-342)已显示与抗体介导的避孕有关。在这项研究中,我们调查了工厂中mZP3的产量,作为口腔适用的宿主,并在BALB/c小鼠模型中检测了该小肽的免疫原性。将mZP3肽插入乙型肝炎核心抗原的主要免疫显性区域,并通过农杆菌介导的瞬时表达在本氏烟草植物中产生。可溶性HBcAg-mZP3的积累水平高达2.63mg/g叶片干重(LDW),含有〜172µg/mgLDWmZP3肽。蔗糖梯度分析和电子显微镜显示了可溶性蛋白部分中HBcAg-mZP3病毒样颗粒(VLP)的组装。皮下施用在HBcAgVLP上展示的mZP3肽在相对低剂量(每剂量5.5µgmZP3)的BALB/c小鼠中具有免疫原性,并导致产生与野生小鼠的天然透明带结合的mZP3特异性抗体。口服表达HBcAg-mZP3的干叶也引发了与野生小鼠透明带交叉反应的mZP3特异性血清IgG和粘膜IgA。根据这些结果,值得研究植物产生HBcAg-mZP3VLP作为免疫原性可食用诱饵的效率,通过诱导与透明带交叉反应的抗体来降低野生小鼠的生育力。