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Production of monoclonal antibodies against 53-kDa protein of Porphyromonas gingivalis in transgenic rice cell suspension culture

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Abstract

Periodontitis is a bacterial infection-induced inflammatory disease that affects the tissues surrounding the teeth and is the main cause of tooth loss in adults. Porphyromonas gingivalis, a black-pigmented gram-negative anaerobe, is a major causative agent of periodontitis. A 53-kDa protein of the bacterium has been demonstrated to be highly immunogenic and recognized by sera from most periodontitis patients, and it is now considered as a potential target of protective immunity. In this study, complementary DNA encoding monoclonal antibodies to 53-kDa protein of P. gingivalis A7A1-28 was cloned from hybridoma cell lines expressing the antibodies. The heavy and light chain genes were constructed into plant expression vectors under the control of the rice α-amylase 3D (RAmy3D) promoter. The recombinant plant expression vectors were introduced into rice cells (Oryza sativa L. cv. Dongjin) using particle bombardment transformation method. Transgenic rice calli were selected by genomic DNA PCR and used to establish cell suspension cultures. The expression of the heavy and light chain genes of the monoclonal antibodies in the rice cell suspension cultures was detected by western blot analysis. The plant-produced monoclonal antibodies showed specific reactivity with 53-kDa proteins of P. gingivalis. These results suggest that 53-kDa protein-specific monoclonal antibodies produced in the rice cell suspension cultures are biologically active against the bacterial protein and can be used for passive immunization to prevent P. gingivalis-induced periodontal disease.

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References

  • Arai M, Hamada N, Umemoto T (2000) Purification and characterization of a novel secondary fimbrial protein from Porphyromonas gingivalis strain 381. FEMS Microbiol Lett 193:75–81

    Article  CAS  PubMed  Google Scholar 

  • Casadevall A (2002) Passive antibody administration (immediate immunity) as a specific defense against biological weapons. Emerg Infect Dis 8:833–841

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen PW, Lu CA, Yu TS et al (2002) Rice alpha-amylase transcriptional enhancers direct multiple mode regulation of promoters in transgenic rice. J Biol Chem 277:13641–13649

    Article  CAS  PubMed  Google Scholar 

  • Clifano JV, Gunsolley JC, Schenkein HA et al (1997) A comparison of IgG antibody reactive with Bacteroides forsythus and Porphyromonas gingivalis in adult and early-onset periodontitis. J Periodontol 68:734–738

    Article  Google Scholar 

  • De Neve M, De Loose M, Jacobs A et al (1993) Assembly of an antibody and its derived antibody fragment in Nicotiana and Arabidopsis. Transgenic Res 2:227–237

    Article  PubMed  Google Scholar 

  • Enersen M, Nakano K, Amano A (2013) Porphyromonas gingivalis fimbriae. J Oral Microbiol 5:20265

    Article  Google Scholar 

  • Giritch A, Marillonnet S, Engler C et al (2006) Rapid-high yield expression of full-size IgG antibodies in plants coinfiltrated with noncompeting viral vectors. Proc Natl Acad Sci USA 103:14701–14706

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gleba Y, Klimyuk V, Marillonnet S (2005) Magnifection a new platform for expressing recombinant vaccines in plants. Vaccine 23:2042–2048

    Article  CAS  PubMed  Google Scholar 

  • Guo S, Takahashi K, Kokeguchi S et al (2000) Antibody responses against Porphyromonas gingivalis infection in patients with early-onset periodontitis. J Clin Periodontol 27:769–777

    Article  CAS  PubMed  Google Scholar 

  • Hajishengallis G (2015) Periodontitis: from microbial immune subversion to systemic inflammation. Nat Rev Immunol 15:30–44

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hamada N, Watanabe K, Sasakawa C et al (1994) Construction and characterization of a fimA mutant of Porphyromonas gingivalis. Infect Immun 62:1696–1704

    CAS  PubMed  PubMed Central  Google Scholar 

  • Hamada N, Sojar HT, Cho MI et al (1996) Isolation and characterization of a minor fimbria from Porphyromonas gingivalis. Infect Immun 64:4788–4794

    CAS  PubMed  PubMed Central  Google Scholar 

  • Hiatt A, Cafferkey R, Bowdish K (1989) Production of antibodies in transgenic plants. Nature 342:76–78

    Article  CAS  PubMed  Google Scholar 

  • Huang TK, McDonald KA (2012) Molecular farming using bioreactor-based plant cell suspension cultures for recombinant protein production. In: Wang A, Ma S (eds) Molecular farming in plants: recent advances and future prospects. Springer, Dordrecht, pp 37–67

    Chapter  Google Scholar 

  • Huang Z, Phoolcharoen W, Lai H et al (2010) High-level rapid production of full-size monoclonal antibodies in plants by a single-vector DNA replicon system. Biotechnol Bioeng 106:9–17

    CAS  PubMed  PubMed Central  Google Scholar 

  • Iwata T, Yamato M, Ishikawa I et al (2014) Tissue engineering in periodontal tissue. Anat Rec (Hoboken) 297:16–25

    Article  CAS  Google Scholar 

  • Kim J, Amar S (2006) Periodontal disease and systemic conditions: a bidirectional relationship. Odontology 94:10–21

    Article  PubMed  PubMed Central  Google Scholar 

  • Kim MY, Yang MS, Kim TG (2012a) Expression of a consensus dengue virus envelope protein domain III in transgenic callus of rice. Plant Cell Tiss Organ Cult 109:509–515

    Article  CAS  Google Scholar 

  • Kim TG, Kim BG, Jeong DK et al (2012b) Production of monoclonal antibodies against the FimA protein of Porphyromonas gingivalis in Nicotiana benthamiana. Biotechnol Bioprocess Eng 17:420–426

    Article  CAS  Google Scholar 

  • Kim NS, Jang SH, Yu HY et al (2013) Amylase and cysteine proteinase gene knockdown in rice cells using RNA interference for enhancing production of recombinant proteins. Plant Cell Tiss Organ Cult 114:97–107

    Article  CAS  Google Scholar 

  • Kim BG, Kim SH, Kim NS et al (2014) Production of monoclonal antibody against FimA protein from Porphyromonas gingivalis in rice cell suspension culture. Plant Cell Tiss Organ Cult 118:293–304

    Article  CAS  Google Scholar 

  • Koh EM, Kim J, Kim TG et al (2011) Cloning and characterization of heavy and light chain genes encoding the FimA-specific monoclonal antibodies that inhibit Porphyromonas gingivalis adhesion. Microbiol Immunol 55:199–210

    Article  CAS  PubMed  Google Scholar 

  • Kokeguchi S, Kato K, Kurihara H et al (1990) Purification and characterization of two major outer membrane proteins from Porphyromonas gingivalis. Dent Jpn (Tokyo) 27:29–34

    CAS  Google Scholar 

  • Kolotilin I, Topp E, Cox E et al (2014) Plant-based solutions for veterinary immunotherapeutics and prophylactics. Vet Res 45:117

    Article  PubMed  Google Scholar 

  • Kurihara H, Nishimura F, Nakamura T et al (1991) Humoral immune response to an antigen from Porphyromonas gingivalis 381 in periodontal disease. Infect Immun 59:2758–2762

    CAS  PubMed  PubMed Central  Google Scholar 

  • Lamont RJ, Jenkinson HF (1998) Life below the gum line: pathogenic mechanisms of Porphyromonas gingivalis. Microbiol Mol Biol Rev 62:1244–1263

    CAS  PubMed Central  Google Scholar 

  • Larrick JW, Yu L, Naftzger C et al (2001) Production of secretory IgA antibodies in plants. Biomol Eng 18:87–94

    Article  CAS  PubMed  Google Scholar 

  • Loesche WJ, Grossman NS (2001) Periodontal disease as a specific, albeit chronic, infection: diagnosis and treatment. Microbiol Rev 14:727–752

    Article  CAS  Google Scholar 

  • Ma JK, Hiatt A, Hein M et al (1995) Generation and assembly of secretory antibodies in plants. Science 268:716–719

    Article  CAS  PubMed  Google Scholar 

  • Ma JK, Hikmat BY, Wycoff K et al (1998) Characterization of a recombinant plant monoclonal secretory antibody and preventive immunotherapy in humans. Nat Med 4:601–606

    Article  CAS  PubMed  Google Scholar 

  • Machtei EE, Dunford R, Hausmann E et al (1997) Longitudinal study of prognostic factors in established periodontitis patients. J Clin Periodontol 24:102–109

    Article  CAS  PubMed  Google Scholar 

  • Nagano K, Hasegawa Y, Yoshida Y et al (2015) A major fimbrilin variant of Mfa1 fimbriae in Porphyromonas gingivalis. J Dent Res 94:1143–1148

    Article  CAS  PubMed  Google Scholar 

  • Orzáez D, Granell A, Blázquez MA (2009) Manufacturing antibodies in the plant cell. Biotechnol J 4:1712–1724

    Article  PubMed  Google Scholar 

  • Saini R, Marawar PP, Shete S et al (2009) Periodontitis, a true infection. J Glob Infect Dis 1:149–150

    Article  PubMed  PubMed Central  Google Scholar 

  • Schouten A, Roosien J, De Boer JM et al (1997) Improving scFv antibody expression levels in the plant cytosol. FEBS Lett 415:235–241

    Article  CAS  PubMed  Google Scholar 

  • Shin YJ, Hong SY, Kwon TH et al (2003) High level of expression of recombinant human granulocyte-macrophage colony stimulating factor in transgenic rice cell suspension culture. Biotechnol Bioeng 82:778–783

    Article  CAS  PubMed  Google Scholar 

  • Strugnell R, Zepp F, Cunningham A et al (2011) Vaccine antigens. In: Garçon N, Stern PL, Cunningham AL (eds) Understanding modern vaccines: perspectives in vaccinology. Elsevier, Amsterdam

    Google Scholar 

  • Tavladoraki P, Benvenuto E, Trinca S et al (1993) Transgenic plants expressing a functional single-chain Fv antibody are specifically protected from virus attack. Nature 366:469–472

    Article  CAS  PubMed  Google Scholar 

  • Thompson JA, Abdullah R, Cocking EC (1986) Protoplast culture of rice (Oryza sativa L.) using media solidified with agarose. Plant Sci 47:123–133

    Article  Google Scholar 

  • Vigil A, Davies DH, Felgner PL (2010) Defining the humoral immune response to infectious agents using high-density protein microarrays. Future Microbiol 5:241–251

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zeitlin L, Olmsted SS, Moench TR et al (1998) A humanized monoclonal antibody produced in transgenic plants for immunoprotection of the vagina against genital herpes. Nat Biotechnol 16:1361–1364

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This study was supported by a grant of the Korean Health Technology R&D Project, Ministry of Health and Welfare (HI13C1945) and research funds of Chonbuk National University in 2014.

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Authors and Affiliations

  1. Center for Jeongup Industry-Academy-Institute Cooperation, Chonbuk National University, Jeonju, 54896, Republic of Korea

    Tae-Geum Kim & Moon-Sik Yang

  2. Research Center of Bioactive Materials, Chonbuk National University, Jeonju, 54896, Republic of Korea

    Tae-Geum Kim & Moon-Sik Yang

  3. Department of Bioactive Material Sciences, Chonbuk National University, Jeonju, 54896, Republic of Korea

    Nguyen-Quang-Duc Tien & Moon-Sik Yang

  4. Department of Molecular Biology, Institute of Molecular Biology and Genetics, Chonbuk National University, Jeonju, 54896, Republic of Korea

    Moon-Sik Yang

  5. Department of Maxillofacial Biomedical Engineering, Kyung Hee University, Seoul, 02447, Republic of Korea

    Ji-Hoi Moon & Jin-Yong Lee

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  1. Tae-Geum Kim
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  2. Nguyen-Quang-Duc Tien
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  3. Moon-Sik Yang
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  4. Ji-Hoi Moon
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Correspondence to Jin-Yong Lee.

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Kim, TG., Tien, NQD., Yang, MS. et al. Production of monoclonal antibodies against 53-kDa protein of Porphyromonas gingivalis in transgenic rice cell suspension culture. Plant Cell Tiss Organ Cult 126, 387–397 (2016). https://doi.org/10.1007/s11240-016-1005-6

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  • DOI: https://doi.org/10.1007/s11240-016-1005-6

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