Abstract:
Rodent population control through contraception requires species-specific oral contraceptive vaccines. Therefore, in this study, we produced putative mouse-specific contraceptive peptides, mZP2 (from oocyte) and mIzumo1 (from sperm), in plants using Agrobacterium-mediated transient expression. Peptides were produced separately in Nicotiana benthamiana using constructs encoding antigens containing three copies of each peptide. We also determined the immunogenicity and contraceptive effects of the plant-produced antigens in female BALB/c mice. Mice immunized subcutaneously with a relatively low amount of antigen (5 µg/dose of each peptide in a mixture) showed systemic immune responses against mZP2-3 and mIzumo1-3 antigens. Moreover, the mean litter size of mice treated with the plant-produced antigens was reduced by 39% compared to that of the control mice. Notably, there was a significant negative correlation between the number of pups born and individual antibody levels against both antigens. Immunofluorescence assays demonstrated the binding of induced antibodies to the oocytes of BALB/c and wild-type mice in vivo and in vitro, respectively. Our study demonstrate the feasibility of producing small contraceptive peptides in plants that can be further used to develop oral contraceptive vaccines against mouse populations.
摘要:
通过避孕控制啮齿动物种群需要特定物种的口服避孕疫苗。因此,在这项研究中,我们生产了假定的小鼠特异性避孕肽,mZP2(来自卵母细胞)和mIzumo1(来自精子),在植物中使用农杆菌介导的瞬时表达。使用编码含有每种肽的三个拷贝的抗原的构建体在Nicotianabenthamiana中分别产生肽。我们还确定了植物产生的抗原在雌性BALB/c小鼠中的免疫原性和避孕作用。用相对低量的抗原(5μg/剂量的混合物中的每种肽)皮下免疫的小鼠显示针对mZP2-3和mIzumo1-3抗原的全身免疫应答。此外,与对照小鼠相比,用植物产生的抗原处理的小鼠的平均产仔数减少了39%。值得注意的是,出生的幼崽数量与针对两种抗原的个体抗体水平之间存在显着负相关。免疫荧光实验证明了诱导抗体在体内和体外与BALB/c和野生型小鼠的卵母细胞结合,分别。我们的研究证明了在植物中生产小避孕肽的可行性,这些肽可进一步用于开发针对小鼠群体的口服避孕疫苗。