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조회238

중앙대학교 의과대학

Plant Biotechnol. J., Dec 26 2024, | https://doi.org/10.1111/pbi.14552 Modification of Fc-fusion protein structures to enhance efficacy of cancer vaccine in plant expression system

Sohee Lim¹, Hyun Joo Chung², Yoo Jin Oh³, Peter Hinterdorfer³, Soon Chul Myung², Young-Jin Seo⁴ and Kisung Ko^1,*

¹BioSystems Design Lab, Department of Medicine, College of Medicine, Chung- Ang University, Seoul, Korea
²Department of Urology, College of Medicine, Chung- Ang University, Seoul, Korea
³Department of Applied Experimental Biophysics, Johannes Kepler University, Linz, Austria

⁴Department of Life Science, Chung- Ang University, Seoul, Korea

^*Correspondence : Kisung Ko

Abstract

Epithelial cell adhesion molecule (EpCAM) fused to IgG, IgA and IgM Fc domains was expressed to create IgG, IgA and IgM-like structures as anti-cancer vaccines in Nicotiana tabacum. High-mannose glycan structures were generated by adding a C-terminal endoplasmic reticulum (ER) retention motif (KDEL) to the Fc domain (FcK) to produce EpCAM-Fc and EpCAM-FcK proteins in transgenic plants via Agrobacterium-mediated transformation. Cross-fertilization of EpCAM-Fc (FcK) transgenic plants with Joining chain (J-chain, J and JK) transgenic plants led to stable expression of large quaternary EpCAM-IgA Fc (EpCAM-A) and IgM-like (EpCAM-M) proteins. Immunoblotting, SDS-PAGE and ELISA analyses demonstrated that proteins with KDEL had higher expression levels and binding activity to anti-EpCAM IgGs. IgM showed the strongest binding among the fusion proteins, followed by IgA and IgG. Sera from BALB/c mice immunized with these vaccines produced anti-EpCAM IgGs. Flow cytometry indicated that the EpCAM-Fc fusion proteins significantly activated CD8⁺ cytotoxic T cells, CD4⁺ helper T cells and B cells, particularly with EpCAM-FcK^P and EpCAM-Fc^P (FcK^P) × J^P (JK^P). The induced anti-EpCAM IgGs captured human prostate cancer PC-3 and colorectal cancer SW620 cells. Sera from immunized mice inhibited cancer cell proliferation, migration and invasion; down-regulated proliferation markers (PCNA, Ki-67) and epithelial-mesenchymal transition markers (Vimentin); and up-regulated E-cadherin. These findings suggest that N. tabacum can produce effective vaccine candidates to induce anti-cancer immune responses.

논문정보

형식Research article
게재일2024년 12월 (BRIC 등록일 2024-12-27)
연구진국내(교신)+국외 연구진
분야 생명과학 > 식물학

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