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Point mutations of single amino acids abolish ability of alpha(3) NC1 domain to elicit experimental autoimmune glomerulonephritis in rats

Hellmark, Thomas LU ; Chen, LL; Ohlsson, Sophie LU ; Wieslander, R and Bolton, WK (2003) In Journal of Biological Chemistry 278(47). p.46516-46522
Abstract
We previously showed concordance between Goodpasture syndrome antibody binding and production of experimental glomerulonephritis using human chimeric proteins. We now examine a more limited aminoterminal region of alpha(3)( IV) non- collagenous domain ( NC1) and the impact of single amino acid ( AA) mutations of this region on glomerulonephritis induction. Rats were immunized with collagenase- solubilized glomerular basement membrane ( csGBM), D3, an alpha(1)( IV) NC1 chimeric protein with 69 AA of alpha(3)( IV) NC1 ( binds Goodpasture sera), D4, the D3 construct shortened by 4 AA ( nonbinding), P9, P10, single AA mutants ( non- binding), and S2, alpha(1)( IV) NC1 with 9 AA of alpha(3)( IV) NC1 ( binding). All rats immunized with csGBM and... (More)
We previously showed concordance between Goodpasture syndrome antibody binding and production of experimental glomerulonephritis using human chimeric proteins. We now examine a more limited aminoterminal region of alpha(3)( IV) non- collagenous domain ( NC1) and the impact of single amino acid ( AA) mutations of this region on glomerulonephritis induction. Rats were immunized with collagenase- solubilized glomerular basement membrane ( csGBM), D3, an alpha(1)( IV) NC1 chimeric protein with 69 AA of alpha(3)( IV) NC1 ( binds Goodpasture sera), D4, the D3 construct shortened by 4 AA ( nonbinding), P9, P10, single AA mutants ( non- binding), and S2, alpha(1)( IV) NC1 with 9 AA of alpha(3)( IV) NC1 ( binding). All rats immunized with csGBM and S2 and 50% of D3 rats developed glomerulonephritis. csGBM rats had intense GBM- bound IgG deposits, but S2 and D3 rats had minimal deposits. None of the D4, P9, or P10 rats developed glomerulonephritis. Lymphocytes from nephritic rats proliferated with csGBM, S2, and D3, but not with D4, P9, or P10. Discrete segments of alpha(3)( IV) NC1 within the alpha(1)( IV) NC1 backbone can induce glomerulonephritis. Single AA mutations within that epitope render the antigen unresponsive to Goodpasture sera and incapable of inducing glomerulonephritis. These studies support the concordance of glomerulonephritis inductivity and Goodpasture serum binding. Further, they define a critical limited AA sequence within alpha(3)( IV) NC1 of nine or fewer AA, which confers nephritogenicity to the non-nephritogenic alpha(1)( IV) NC1 without in vivo antibody binding. This region may be a T- cell epitope responsible for induction of glomerulonephritis in this model in rats and Goodpasture syndrome in man. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Biological Chemistry
volume
278
issue
47
pages
46516 - 46522
publisher
ASBMB
external identifiers
  • wos:000186569400036
  • scopus:0344875580
ISSN
1083-351X
DOI
10.1074/jbc.M211951200
language
English
LU publication?
yes
id
8a95bc68-7975-48a3-96c8-582d670d72b7 (old id 295041)
date added to LUP
2007-09-13 08:06:19
date last changed
2018-10-03 10:33:45
@article{8a95bc68-7975-48a3-96c8-582d670d72b7,
  abstract     = {We previously showed concordance between Goodpasture syndrome antibody binding and production of experimental glomerulonephritis using human chimeric proteins. We now examine a more limited aminoterminal region of alpha(3)( IV) non- collagenous domain ( NC1) and the impact of single amino acid ( AA) mutations of this region on glomerulonephritis induction. Rats were immunized with collagenase- solubilized glomerular basement membrane ( csGBM), D3, an alpha(1)( IV) NC1 chimeric protein with 69 AA of alpha(3)( IV) NC1 ( binds Goodpasture sera), D4, the D3 construct shortened by 4 AA ( nonbinding), P9, P10, single AA mutants ( non- binding), and S2, alpha(1)( IV) NC1 with 9 AA of alpha(3)( IV) NC1 ( binding). All rats immunized with csGBM and S2 and 50% of D3 rats developed glomerulonephritis. csGBM rats had intense GBM- bound IgG deposits, but S2 and D3 rats had minimal deposits. None of the D4, P9, or P10 rats developed glomerulonephritis. Lymphocytes from nephritic rats proliferated with csGBM, S2, and D3, but not with D4, P9, or P10. Discrete segments of alpha(3)( IV) NC1 within the alpha(1)( IV) NC1 backbone can induce glomerulonephritis. Single AA mutations within that epitope render the antigen unresponsive to Goodpasture sera and incapable of inducing glomerulonephritis. These studies support the concordance of glomerulonephritis inductivity and Goodpasture serum binding. Further, they define a critical limited AA sequence within alpha(3)( IV) NC1 of nine or fewer AA, which confers nephritogenicity to the non-nephritogenic alpha(1)( IV) NC1 without in vivo antibody binding. This region may be a T- cell epitope responsible for induction of glomerulonephritis in this model in rats and Goodpasture syndrome in man.},
  author       = {Hellmark, Thomas and Chen, LL and Ohlsson, Sophie and Wieslander, R and Bolton, WK},
  issn         = {1083-351X},
  language     = {eng},
  number       = {47},
  pages        = {46516--46522},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Point mutations of single amino acids abolish ability of alpha(3) NC1 domain to elicit experimental autoimmune glomerulonephritis in rats},
  url          = {http://dx.doi.org/10.1074/jbc.M211951200},
  volume       = {278},
  year         = {2003},
}