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Journal of Endocrinological Investigation

, Volume 26, Issue 11, pp 1076–1080 | Cite as

Recombinant monoclonal thyrotropin-stimulation blocking antibody (TSBAb) established from peripheral lymphocytes of a hypothyroid patient with primary myxedema

  • K. Moriyama
  • J. Okuda
  • M. Saijo
  • Y. Hattori
  • N. Kanamoto
  • Y. Hataya
  • F. Matsuda
  • T. Mori
  • K. Nakao
  • Takashi Akamizu
Original Article

Abstract

Anti-TSH receptor antibodies (TRAbs) have been known to be involved in Graves’ disease and primary hypothyroidism. We previously isolated and reconstituted immunoglobulin (Ig) genes of Epstein-Barr virus-transformed B cell clones producing monoclonal TRAbs obtained from Graves’ patients. In the present study, we performed a similar experiment using a B cell clone, 32A-5, derived from a patient with primary hypothyroidism. The variable region genes of Ig heavy (H) and light (L) chains were isolated and sequenced from the 32A-5 clone. A significant number of somatic mutations were found in variable regions of H and L chain gene segments. Each pair of H and L chain cDNAs was ligated into an expression vector for IgG1 production and stably introduced into myeloma cells. The transfectants were injected ip into BALB/c mice to yield ample volume of the antibody for following applications. Interactions of recombinant 32A-5 with Graves’ sera with varying thyroid-stimulating antibody (TSAb) activities were studied. The recombinant antibody tended to suppress TSAb activities in 10 of 15 Graves’ sera, in which four were significantly inhibited. In summary, this is the first study to analyze human monoclonal TSH-stimulation blocking antibodies (TSBAb) at the molecular level. Use of human recombinant monoclonal TSBAb may be an analytical tool for molecular-basis etiology and an alternative therapeutic path for Graves’ disease.

Keywords

Immunoglobulin Heavy Chain Diversity TSAb Activity Primary Myxedema Germline Segment Suppress Thyroid Hormone 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Italian Society of Endocrinology (SIE) 2003

Authors and Affiliations

  • K. Moriyama
    • 1
  • J. Okuda
    • 2
  • M. Saijo
    • 1
  • Y. Hattori
    • 1
  • N. Kanamoto
    • 1
  • Y. Hataya
    • 1
  • F. Matsuda
    • 3
  • T. Mori
    • 2
  • K. Nakao
    • 1
  • Takashi Akamizu
    • 1
    • 2
  1. 1.Department of Medicine and Clinical ScienceKyoto University Graduate School of MedicineKyotoJapan
  2. 2.Department of Laboratory MedicineKyoto University Graduate School of MedicineKyotoJapan
  3. 3.Molecular BiologyKyoto University Graduate School of MedicineKyotoJapan

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