Kang Yangjoo, Kim Deuk-Su, Hwang Hyunjoo, Kim Yerin, Seo Young-Jin, Hinterdorfer Peter, Ko Kisung
Department of Medicine, Medicine of College, Chung-Ang University, Seoul, South Korea.
Department of Life Science, Chung-Ang University, Seoul, South Korea.
Transgenic Res. 2025 Mar 26;34(1):16. doi: 10.1007/s11248-025-00433-0.
Prostatic acid phosphatase (PAP) is a specific protein that is highly expressed in prostate cancer. In this study, we constructed two recombinant PAP fusion genes: PAP fused to the immunoglobulin G (IgG) Fc fragment (designated PAP-Fc) and PAP-Fc fused to the endoplasmic reticulum retention sequence KDEL (designated PAP-FcK). Transgenic Nicotiana tabacum plants expressing these recombinant macromolecular proteins (MPs) were generated using Agrobacterium-mediated transformation, and the presence of both genes was confirmed through genomic PCR. Western blot analysis validated the expression of PAP-Fc and PAP-FcK MPs, which were successfully purified via protein A affinity chromatography. Size-exclusion high-performance liquid chromatography revealed dimeric peaks for PAP-Fc (PAP-Fc) and PAP-FcK (PAP-FcK). Bio-transmission electron microscopy demonstrated 'Y'-shaped protein particles resembling antibody structures. Moreover, PAP-Fc and PAP-FcK exhibited a high association rate with human FcγR and FcRn. Vaccination of mice with both PAP-Fc and PAP-FcK resulted in increased total IgG against PAP and enhanced activation of CD4 T cells, comparable to mice immunized with PAP, which served as a positive control. These findings indicate that both plant-derived MPs can effectively induce adaptive immunity, positioning them as promising candidates for prostate cancer vaccines. Overall, plants expressing PAP-Fc and PAP-FcK represent a viable production system for antigenic macromolecule-based prostate cancer vaccines.
前列腺酸性磷酸酶（PAP）是一种在前列腺癌中高表达的特异性蛋白质。在本研究中，我们构建了两个重组PAP融合基因：与免疫球蛋白G（IgG）Fc片段融合的PAP（命名为PAP-Fc）以及与内质网滞留序列KDEL融合的PAP-Fc（命名为PAP-FcK）。利用农杆菌介导的转化方法培育出表达这些重组大分子蛋白（MPs）的转基因烟草植株，并通过基因组PCR确认了这两个基因的存在。蛋白质免疫印迹分析验证了PAP-Fc和PAP-FcK MPs的表达，它们通过蛋白A亲和层析成功纯化。尺寸排阻高效液相色谱显示PAP-Fc（PAP-Fc）和PAP-FcK（PAP-FcK）出现二聚体峰。生物透射电子显微镜显示出类似抗体结构的“Y”形蛋白颗粒。此外，PAP-Fc和PAP-FcK与人FcγR和FcRn的结合率很高。用PAP-Fc和PAP-FcK对小鼠进行免疫接种后，针对PAP的总IgG增加，CD4 T细胞的活化增强，这与作为阳性对照的用PAP免疫的小鼠相当。这些发现表明，两种植物源MPs都能有效诱导适应性免疫，使其成为前列腺癌疫苗的有希望的候选物。总体而言，表达PAP-Fc和PAP-FcK的植物代表了一种可行的基于抗原大分子的前列腺癌疫苗生产系统。
