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Co-transient expression of PSA-Fc and PAP-Fc fusion protein in plant as prostate cancer vaccine candidates and immune responses in mice

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Abstract

Key message

PAP-FcK and PSA-FcK prostate cancer antigenic proteins transiently co-expressed in plant induce their specific humoral immune responses in mice.

Abstract

Prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP) have been considered as immunotherapeutic antigens for prostate cancer. The use of a single antigenic agent is unlikely to be effective in eliciting immunotherapeutic responses due to the heterogeneous and multifocal nature of prostate cancer. Thus, multiple antigens have been combined to enhance their anti-cancer effects. In the current study, PSA and PAP were fused to the crystallizable region (Fc region) of immunoglobulin G1 and tagged with KDEL, the endoplasmic reticulum (ER) retention signal motif, to generate PSA-FcK and PAP-FcK, respectively, and were transiently co-expressed in Nicotiana benthamiana. Western blot analysis confirmed the co-expression of PSA-FcK and PAP-FcK (PSA-FcK + PAP-FcK) with a 1:3 ratios in the co-infiltrated plants. PSA-FcK, PAP-FcK, and PSA-FcK + PAP-FcK proteins were successfully purified from N. benthamiana by protein A affinity chromatography. ELISA showed that anti-PAP and anti-PSA antibodies successfully detected PAP-FcK and PSA-FcK, respectively, and both detected PSA-FcK + PAP-FcK. Surface plasmon resonance (SPR) analysis confirmed the binding affinity of the plant-derived Fc fusion proteins to FcγRI/CD64. Furthermore, we also confirmed that mice injected with PSA-FcK + PAP-FcK produced both PSA- and PAP-specific IgGs, demonstrating their immunogenicity. This study suggested that the transient plant expression system can be applied to produce the dual-antigen Fc fusion protein (PSA-FcK + PAP-FcK) for prostate cancer immunotherapy.

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Funding

This research was supported by the National Research Foundation of Korea grant [2021R1F1A1063869], Bio & Medical Technology Development Program of the National Research Foundation (NRF) and funded by the Korean government (MSIT) (No. 2019K1A3A1A18116087).

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Author notes

  1. Soyeon Oh and Kibum Kim have contributed equally to this work.

Authors and Affiliations

  1. Department of Medical Science, College of Medicine, Chung-Ang University, Seoul, South Korea

    Soyeon Oh, Kibum Kim, Yang Joo Kang, Hyunjoo Hwang, Yerin Kim & Kisung Ko

  2. Institute of Biophysics, Johannes Kepler University, 4020, Linz, Austria

    Peter Hinterdorfer

  3. Department of Pathology, College of Medicine, Chung-Ang University, Seoul, South Korea

    Mi Kyung Kim

  4. Department of Stem Cell Biology, Konkuk University School of Medicine, Konkuk University, Seoul, South Korea

    Kinarm Ko

  5. Institute of Plasma Technology, Korea Institute of Fusion Energy, 37 Dongjangsan-ro, Gunsan-si, 54004, South Korea

    Young Koung Lee

  6. Vegetable Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, Wanju-gun, South Korea

    Do-Sun Kim

  7. Department of Urology, College of Medicine, Chung-Ang University, Seoul, South Korea

    Soon Chul Myung

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  1. Soyeon Oh
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Contributions

Conceptualization, SYO, and KK; methodology, SYO, KBK, YJK, HJH, YRK, and PH; validation, MKK, YKL, DSK, and SCM.; formal analysis, SYO; investigation, KBK, and KK; resources, SYO, PH, and KK; data curation, KBK and YJK; writing—original draft preparation, SYO and KBK; writing—review and editing, KK; visualization, KBK and KK; supervision, KK; project administration, KBK and Y.J.K.

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Correspondence to Kisung Ko.

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Communicated by Baochun Li.

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Oh, S., Kim, K., Kang, Y.J. et al. Co-transient expression of PSA-Fc and PAP-Fc fusion protein in plant as prostate cancer vaccine candidates and immune responses in mice. Plant Cell Rep 42, 1203–1215 (2023). https://doi.org/10.1007/s00299-023-03028-3

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