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In vitro and in vivo expression of a nephritogenic Ig heavy chain determinant: pathogenic autoreactivity requires permissive light chains.

Publication ,  Journal Article
Cooperstone, BG; Rahman, MM; Rudolph, EH; Foster, MH
Published in: Immunol Cell Biol
June 2001

Lymphocyte antigen receptors are promising targets for immune intervention strategies in disorders marked by repertoire skewing or expansion of lymphocyte subsets. Appropriate application of immune receptor modulation is predicated on understanding the role of a particular receptor in pathogenesis and disease regulation. The VHB/W16 gene, restricted to mice carrying the j haplotype for the J558 family, is overexpressed by murine lupus anti-DNA Ig. This gene is also expressed recurrently among nephritogenic anti-DNA Ig recovered from several autoimmune strains, suggesting that cells expressing this pathogenic receptor are positively selected during disease progression. To explore the extent and mechanisms by which Ig H chains expressing this gene contribute to autoimmunity, an Ig H chain gene was engineered for in vitro and in vivo recombination studies. Site-directed mutagenesis generated unique restriction sites to link PCR-amplified V region (VDJ) cDNA to previously isolated genomic fragments containing Ig regulatory and signal sequences. The new 3 kb VDJ gene was then ligated to a 9 kb fragment encoding the IgM constant region. Transfection of H chain loss variant myeloma with the complete 12 kb construct, termed 238H-Cmicro, resulted in secretion of intact Ig pairing 238H-Cmicro, with a lambda L chain; however, transfectant Ig lacked autoreactivity and pathogenicity. Introduction of the 238H-Cmicro H chain as a transgene onto the non-autoimmune C57BL/6 background resulted in abundant B cell surface expression of 238H-Cmicro, however, four transgenic Ig recovered by fusion of LPS-stimulated splenocytes and formed by combination of 238H-Cmicro, with endogenous kappa chains do not bind DNA or laminin. These results indicate that the antigen binding sites encoded by this disease-associated gene and/or H chain must associate with permissive L chains to specify autoimmunity. The 238H-Cmicro, transgenic model should prove useful in dissecting the in vivo fate of 238H-Cmicro, L combinations that produce pathogenic autoreactive receptors and in evaluating receptor-targeted interventions.

Duke Scholars

Published In

Immunol Cell Biol

DOI

ISSN

0818-9641

Publication Date

June 2001

Volume

79

Issue

3

Start / End Page

222 / 230

Location

United States

Related Subject Headings

  • Transfection
  • Sequence Alignment
  • Recombinant Fusion Proteins
  • Molecular Sequence Data
  • Mice, Transgenic
  • Mice, Inbred C57BL
  • Mice, Inbred BALB C
  • Mice
  • Membrane Glycoproteins
  • Immunology
 

Citation

APA
Chicago
ICMJE
MLA
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Cooperstone, B. G., Rahman, M. M., Rudolph, E. H., & Foster, M. H. (2001). In vitro and in vivo expression of a nephritogenic Ig heavy chain determinant: pathogenic autoreactivity requires permissive light chains. Immunol Cell Biol, 79(3), 222–230. https://doi.org/10.1046/j.1440-1711.2001.01001.x
Cooperstone, B. G., M. M. Rahman, E. H. Rudolph, and M. H. Foster. “In vitro and in vivo expression of a nephritogenic Ig heavy chain determinant: pathogenic autoreactivity requires permissive light chains.Immunol Cell Biol 79, no. 3 (June 2001): 222–30. https://doi.org/10.1046/j.1440-1711.2001.01001.x.
Cooperstone BG, Rahman MM, Rudolph EH, Foster MH. In vitro and in vivo expression of a nephritogenic Ig heavy chain determinant: pathogenic autoreactivity requires permissive light chains. Immunol Cell Biol. 2001 Jun;79(3):222–30.
Cooperstone, B. G., et al. “In vitro and in vivo expression of a nephritogenic Ig heavy chain determinant: pathogenic autoreactivity requires permissive light chains.Immunol Cell Biol, vol. 79, no. 3, June 2001, pp. 222–30. Pubmed, doi:10.1046/j.1440-1711.2001.01001.x.
Cooperstone BG, Rahman MM, Rudolph EH, Foster MH. In vitro and in vivo expression of a nephritogenic Ig heavy chain determinant: pathogenic autoreactivity requires permissive light chains. Immunol Cell Biol. 2001 Jun;79(3):222–230.

Published In

Immunol Cell Biol

DOI

ISSN

0818-9641

Publication Date

June 2001

Volume

79

Issue

3

Start / End Page

222 / 230

Location

United States

Related Subject Headings

  • Transfection
  • Sequence Alignment
  • Recombinant Fusion Proteins
  • Molecular Sequence Data
  • Mice, Transgenic
  • Mice, Inbred C57BL
  • Mice, Inbred BALB C
  • Mice
  • Membrane Glycoproteins
  • Immunology