Site-specific antibodies distinguish single amino acid substitutions in position 57 in HLA-DQ β-chain alleles associated with insulin-dependent diabetes
(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
- Atar, D. ; Dyrberg, T. ; Michelsen, B. ; Karlsen, A. LU ; Kofod, H. ; Molvig, J. and Lernmark, A. LU
- publishing date
- 1989-01-01
- 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}}, }