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Recovery of a human natural antibody against the noncollagenous-1 domain of type IV collagen using humanized models.

Publication ,  Journal Article
Worni-Schudel, IM; Clark, AG; Chien, T; Hwang, K-K; Chen, BJ; Foster, MH
Published in: J Transl Med
June 6, 2015

BACKGROUND: Anti-glomerular basement membrane nephritis and Goodpasture syndrome result from autoantibody (Ab)-mediated destruction of kidney and lung. Ab target the noncollagenous 1 (NC1) domain of alpha3(IV) collagen, but little is known about Ab origins or structure. This ignorance is due in part to the inability to recover monoclonal Ab by transformation of patients' blood cells. The aim of this study was to assess the suitability of two humanized models for this purpose. METHODS: NOD-scid-gamma immunodeficient mice were engrafted either with human CD34+ hematopoietic stem cells (HSC) (Hu-HSC mice) and immunized with alpha3(IV)NC1 collagen containing the Goodpasture epitopes or with nephritis patients' peripheral blood leukocytes (PBL) (Hu-PBL mice). After in vivo immune cell development and/or expansion, recovered human B cells were Epstein Barr virus (EBV)-transformed, screened for antigen (Ag) binding, electrofused with a mouse-human heterohybridoma, subcloned, and human Ab RNA sequenced by PCR after reverse transcription to cDNA. Flow cytometry was used to assess human B cell markers and differentiation in Hu-PBL mice. RESULTS: Sequence analysis of a human Ab derived from an immunized Hu-HSC mouse and reactive with alpha3(IV)NC1 collagen reveals that it is encoded by unmutated heavy and light chain genes. The heavy chain complementarity determining region 3, a major determinant of Ag binding, contains uncommon motifs, including an N-region somatically-introduced highly hydrophobic tetrapeptide and dual cysteines encoded by a uniquely human IGHD2-2 Ab gene segment that lacks a murine counterpart. Comparison of human and mouse autoantibodies suggests that structurally similar murine Ab may arise by convergent selection. In contrast to the Hu-HSC model, transformed human B cells are rarely recovered from Hu-PBL mice, in which human B cells terminally differentiate and lose expression of EBV receptor CD21, thus precluding their transformation and recovery. CONCLUSIONS: Hu-HSC mice reveal that potentially pathogenic B cells bearing unmutated Ig receptors reactive with the NC1 domain on alpha3(IV) collagen can be generated in, and not purged from, the human preimmune repertoire. Uniquely human gene elements are recruited to generate the antigen binding site in at least a subset of these autoantibodies, indicating that humanized models may provide insights inaccessible using conventional mouse models.

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Published In

J Transl Med

DOI

EISSN

1479-5876

Publication Date

June 6, 2015

Volume

13

Start / End Page

185

Location

England

Related Subject Headings

  • Tissue Donors
  • Sequence Analysis, Protein
  • Receptors, Complement 3d
  • Protein Structure, Tertiary
  • Nephritis
  • Molecular Sequence Data
  • Models, Animal
  • Mice
  • Lymphocyte Subsets
  • Leukocytes
 

Citation

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Worni-Schudel, I. M., Clark, A. G., Chien, T., Hwang, K.-K., Chen, B. J., & Foster, M. H. (2015). Recovery of a human natural antibody against the noncollagenous-1 domain of type IV collagen using humanized models. J Transl Med, 13, 185. https://doi.org/10.1186/s12967-015-0539-4
Worni-Schudel, Inge M., Amy G. Clark, Tiffany Chien, Kwan-Ki Hwang, Benny J. Chen, and Mary H. Foster. “Recovery of a human natural antibody against the noncollagenous-1 domain of type IV collagen using humanized models.J Transl Med 13 (June 6, 2015): 185. https://doi.org/10.1186/s12967-015-0539-4.
Worni-Schudel IM, Clark AG, Chien T, Hwang K-K, Chen BJ, Foster MH. Recovery of a human natural antibody against the noncollagenous-1 domain of type IV collagen using humanized models. J Transl Med. 2015 Jun 6;13:185.
Worni-Schudel, Inge M., et al. “Recovery of a human natural antibody against the noncollagenous-1 domain of type IV collagen using humanized models.J Transl Med, vol. 13, June 2015, p. 185. Pubmed, doi:10.1186/s12967-015-0539-4.
Worni-Schudel IM, Clark AG, Chien T, Hwang K-K, Chen BJ, Foster MH. Recovery of a human natural antibody against the noncollagenous-1 domain of type IV collagen using humanized models. J Transl Med. 2015 Jun 6;13:185.
Journal cover image

Published In

J Transl Med

DOI

EISSN

1479-5876

Publication Date

June 6, 2015

Volume

13

Start / End Page

185

Location

England

Related Subject Headings

  • Tissue Donors
  • Sequence Analysis, Protein
  • Receptors, Complement 3d
  • Protein Structure, Tertiary
  • Nephritis
  • Molecular Sequence Data
  • Models, Animal
  • Mice
  • Lymphocyte Subsets
  • Leukocytes