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Site-specific antibodies distinguish single amino acid substitutions in position 57 in HLA-DQ β-chain alleles associated with insulin-dependent diabetes

Atar, D. ; Dyrberg, T. ; Michelsen, B. ; Karlsen, A. LU ; Kofod, H. ; Molvig, J. and Lernmark, A. LU orcid (1989) In Journal of Immunology 143(2). p.533-538
Abstract

The HLA-DQ β-chain gene shows a close association with susceptibility or resistance to autoimmune insulin-dependent diabetes mellitus (IDDM) and it has been suggested that the amino acid in position 57 may be of pathogenetic importance. To study the expression of the IDDM associated HLA-DQ β-chain alleles, we immunized rabbits with 12 to 13 amino acid long peptides representing HLA-DQw7 and -DQw8 allelic sequences, differing only by one amino acid in position 57 being aspartic acid (Asp) and alanine (Ala), respectively. Immunoblot analysis of lymphoblastoid cells showed that several antisera recognized a 29-kDa protein, equivalent to the expected molecular size of the HLA-DQ β-chain to yield two antisera for HLA-DQw7 (pos. 57(Asp)) and... (More)

The HLA-DQ β-chain gene shows a close association with susceptibility or resistance to autoimmune insulin-dependent diabetes mellitus (IDDM) and it has been suggested that the amino acid in position 57 may be of pathogenetic importance. To study the expression of the IDDM associated HLA-DQ β-chain alleles, we immunized rabbits with 12 to 13 amino acid long peptides representing HLA-DQw7 and -DQw8 allelic sequences, differing only by one amino acid in position 57 being aspartic acid (Asp) and alanine (Ala), respectively. Immunoblot analysis of lymphoblastoid cells showed that several antisera recognized a 29-kDa protein, equivalent to the expected molecular size of the HLA-DQ β-chain to yield two antisera for HLA-DQw7 (pos. 57(Asp)) and three antisera for HLA-DQw8 (pos. 57(Ala)) positive cells. Analysis of HLA-DR 3/4 positive IDDM patients (n = 24) and controls (n = 19) showed that all (100%) patients were positive for pos. 57(Ala) antiserum compared to 13 of 19 (68%) of the controls. The remaining six controls reacted with the pos. 57(Asp) antisera, whereas none of the patients did. We have therefore successfully been able to generate site-specific antibodies that distinguish single amino acid substitutions in predetermined positions of allelic HLA-DQ β-chain gene products. Such sera should become useful to detect and investigate HLA associated susceptibility to autoimmune diseases in man.

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author
; ; ; ; ; and
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Immunology
volume
143
issue
2
pages
533 - 538
publisher
American Association of Immunologists
external identifiers
  • scopus:0024393505
  • pmid:2738402
ISSN
0022-1767
language
English
LU publication?
no
id
e168f03d-71dd-4bca-aca5-beba086a4ef1
alternative location
https://www.jimmunol.org/content/jimmunol/143/2/533.full.pdf
date added to LUP
2019-09-11 09:58:21
date last changed
2024-03-13 08:14:16
@article{e168f03d-71dd-4bca-aca5-beba086a4ef1,
  abstract     = {{<p>The HLA-DQ β-chain gene shows a close association with susceptibility or resistance to autoimmune insulin-dependent diabetes mellitus (IDDM) and it has been suggested that the amino acid in position 57 may be of pathogenetic importance. To study the expression of the IDDM associated HLA-DQ β-chain alleles, we immunized rabbits with 12 to 13 amino acid long peptides representing HLA-DQw7 and -DQw8 allelic sequences, differing only by one amino acid in position 57 being aspartic acid (Asp) and alanine (Ala), respectively. Immunoblot analysis of lymphoblastoid cells showed that several antisera recognized a 29-kDa protein, equivalent to the expected molecular size of the HLA-DQ β-chain to yield two antisera for HLA-DQw7 (pos. 57(Asp)) and three antisera for HLA-DQw8 (pos. 57(Ala)) positive cells. Analysis of HLA-DR 3/4 positive IDDM patients (n = 24) and controls (n = 19) showed that all (100%) patients were positive for pos. 57(Ala) antiserum compared to 13 of 19 (68%) of the controls. The remaining six controls reacted with the pos. 57(Asp) antisera, whereas none of the patients did. We have therefore successfully been able to generate site-specific antibodies that distinguish single amino acid substitutions in predetermined positions of allelic HLA-DQ β-chain gene products. Such sera should become useful to detect and investigate HLA associated susceptibility to autoimmune diseases in man.</p>}},
  author       = {{Atar, D. and Dyrberg, T. and Michelsen, B. and Karlsen, A. and Kofod, H. and Molvig, J. and Lernmark, A.}},
  issn         = {{0022-1767}},
  language     = {{eng}},
  month        = {{01}},
  number       = {{2}},
  pages        = {{533--538}},
  publisher    = {{American Association of Immunologists}},
  series       = {{Journal of Immunology}},
  title        = {{Site-specific antibodies distinguish single amino acid substitutions in position 57 in HLA-DQ β-chain alleles associated with insulin-dependent diabetes}},
  url          = {{https://www.jimmunol.org/content/jimmunol/143/2/533.full.pdf}},
  volume       = {{143}},
  year         = {{1989}},
}