Generation of high-affinity antibody against T cell-dependent antigen in the Ganp gene-transgenic mouse

Nobuo Sakaguchi, Tetsuya Kimura, Shuzo Matsushita, Satoru Fujimura, Junji Shibata, Masatake Araki, Tamami Sakamoto, Chiemi Minoda, Kazuhiko Kuwahara

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42 Citations (Scopus)

Abstract

Generation of high-affinity Ab is impaired in mice lacking germinal center-associated DNA primase (GANP) in B cells. In this study, we examined the effect of its overexpression in ganp transgenic C57BL/6 mice (Ganp Tg). GanpTg displayed normal phenotype in B cell development, serum Ig levels, and responses against T cell-independent Ag; however, it generated the Ab with much higher affinity against nitrophenyl-chicken gammaglobulin in comparison with C57BL/6. To further examine the affinity increase, we established hybridomas producing high-affinity mAbs and compared their affinities using BIAcore. C57BL/6 generated high-affinity anti-nitrophenyl mAbs (KD ∼ 2.50 ∼ 10-7 M) of IgG1/λ1 and contained the VH186.2 region with W33L mutation. GanpTg generated much higher affinity (KD > 1.57 × 10-9 M) by usage of VH186.2 as well as noncanonical VH7183 regions. GanpTg also generated exceptionally high-affinity anti-HIV-1 (V3 peptide) mAbs (KD > 9.90 × 10-11 M) with neutralizing activity. These results demonstrated that GANP is involved in V region alteration generating high-affinity Ab.

Original languageEnglish
Pages (from-to)4485-4494
Number of pages10
JournalJournal of Immunology
Volume174
Issue number8
DOIs
Publication statusPublished - 15-04-2005

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

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    Sakaguchi, N., Kimura, T., Matsushita, S., Fujimura, S., Shibata, J., Araki, M., Sakamoto, T., Minoda, C., & Kuwahara, K. (2005). Generation of high-affinity antibody against T cell-dependent antigen in the Ganp gene-transgenic mouse. Journal of Immunology, 174(8), 4485-4494. https://doi.org/10.4049/jimmunol.174.8.4485