Skip to main content

Advertisement

Log in

In vitro immunization of Epstein–Barr virus-immortalized B cells augments antigen-specific antibody production

  • JAACT Special Issue
  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

The current method for in vitro immunization (IVI) uses several antigens including toxins, food allergens, pathogenic bacteria, and self-antigen-derived peptides that induce an antigen-specific immune response in peripheral blood mononuclear cells (PBMCs). This protocol, however, requires donor blood collection and preparation of PBMCs before every IVI. In the present study, we aimed to design a more efficient system utilizing B cells immortalized with Epstein–Barr virus (EBV-B) as host cells for IVI to make antigen-specific antibodies. Results showed that previously antigen-sensitized, EBV-B cells exposed to the antigen along with IL-6, CpG oligonucleotides, and CD40 ligand signal produced antigen-specific antibodies. These results provide evidence for a novel and easy method to expand memory-type B cells and produce antigen-specific antibodies.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Brezinsky SC, Chiang GG, Szilvasi A, Mohan S, Shapiro RI, MacLean A, Sisk W, Thill G (2003) A simple method for enriching populations of transfected CHO cells for cells of higher specific productivity. J Immunol Methods 277:141–155

    Google Scholar 

  • Co MS, Queen C (1991) Humanized antibodies for therapy. Nature 351:501–502. doi:10.1038/351501a0

    Article  CAS  Google Scholar 

  • Fujiki T, Tsuji A, Matsumoto SE, Yamashita M, Teruya K, Shirahata S, Katakura Y (2010) Generation of a human anti-tumor necrosis factor-α monoclonal antibody by in vitro immunization with a multiple antigen peptide. Biosci Biotechnol Biochem 74:1836–1840

    Google Scholar 

  • Geylis V, Steinitz M (2006) Immunotherapy of Alzheimer’s disease (AD): from murine models to anti-amyloid beta (Abeta) human monoclonal antibodies. Autoimmun Rev 5:33–39. doi:10.1016/j.autrev.2005.06.007

    Article  Google Scholar 

  • Ichikawa A, Katakura Y, Teruya K, Hashizume S, Shirahata S (1999) In vitro immunization of human peripheral blood lymphocytes: establishment of B cell lines secreting IgM specific for cholera toxin B subunit from lymphocytes stimulated with IL-2 and IL-4. Cytotechnology 31:133–341

    Google Scholar 

  • Katakura Y, Seto P, Miura T, Ohashi H, Teruya K, Shirahata S (1999) Productivity enhancement of recombinant protein in CHO cells via specific promoter activation by oncogenes. Cytotechnology 31:103–109

    Google Scholar 

  • Oxenius A, Campbell KA, Maliszewski CR, Kishimoto T, Kikutani H, Hengartner H, Zinkernagel RM, Bachmann MF (1996) CD40–CD40 ligand interactions are critical in T-B cooperation but not for other anti-viral CD4+ T cell functions. J Exp Med 183:2209–2218

    Google Scholar 

  • Shim SY, Katakura Y, Ichikawa A, Teruya K, Matsuda T, Shirahata S (2001) Epitope analysis of human monoclonal antibody specific for rice allergenic protein generated by in vitro immunization. Cytotechnology 36:109–115. doi:10.1023/A:1014012214095

    Google Scholar 

  • Tamura T, Tomimatsu K, Katakura Y, Yamashita M, Matsumoto SE, Aiba Y, Jung YS, Abe Y, Fujiki T, Teruya K, Shirahata S (2007) Anti-peptide antibody production elicited by in vitro immunization of human peripheral blood mononuclear cells. Biosci Biotechnol Biochem 71:2871–2875

    Google Scholar 

  • Verthelyi D, Ishii KJ, Gursel M, Takeshita F, Klinman DM (2001) Human peripheral blood cells differentially recognize and respond to two distinct CPG motifs. J Immunol 166:2372–2377

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Yoshinori Katakura.

Additional information

Gakuro Harada and Shin-ei Matsumoto corresponded equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Harada, G., Matsumoto, Se., Yamashita, M. et al. In vitro immunization of Epstein–Barr virus-immortalized B cells augments antigen-specific antibody production. Cytotechnology 65, 979–983 (2013). https://doi.org/10.1007/s10616-013-9596-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10616-013-9596-9

Keywords

Navigation