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

D Atar, T Dyrberg, B Michelsen, A Karlsen, H Kofod, J Mølvig and A Lernmark
J Immunol July 15, 1989, 143 (2) 533-538;
D Atar
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T Dyrberg
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B Michelsen
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A Karlsen
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H Kofod
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J Mølvig
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A Lernmark
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Abstract

The HLA-DQ beta-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 beta-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 beta-chain to yield two antisera specific for HLA-DQw7 (pos. 57Asp) and three antisera for HLA-DQw8 (pos. 57Ala) 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. 57Ala antiserum compared to 13 of 19 (68%) of the controls. The remaining six controls reacted with the pos. 57Asp 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 beta-chain gene products. Such sera should become useful to detect and investigate HLA associated susceptibility to autoimmune diseases in man.

  • Copyright © 1989 by American Association of Immunologists

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The Journal of Immunology
Vol. 143, Issue 2
15 Jul 1989
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Site-specific antibodies distinguish single amino acid substitutions in position 57 in HLA-DQ beta-chain alleles associated with insulin-dependent diabetes.
D Atar, T Dyrberg, B Michelsen, A Karlsen, H Kofod, J Mølvig, A Lernmark
The Journal of Immunology July 15, 1989, 143 (2) 533-538;

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Site-specific antibodies distinguish single amino acid substitutions in position 57 in HLA-DQ beta-chain alleles associated with insulin-dependent diabetes.
D Atar, T Dyrberg, B Michelsen, A Karlsen, H Kofod, J Mølvig, A Lernmark
The Journal of Immunology July 15, 1989, 143 (2) 533-538;
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Print ISSN 0022-1767        Online ISSN 1550-6606