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

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Site-specific antibodies distinguish single amino acid substitutions in position 57 in HLA-DQ beta-chain alleles associated with insulin-dependent diabetes. / Atar, D; Dyrberg, T; Michelsen, Birgitte; Karlsen, Alan E; Kofod, Hans; Mølvig, J; Lernmark, A.

In: Journal of Immunology, Vol. 143, No. 2, 15.07.1989, p. 533-8.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Atar, D, Dyrberg, T, Michelsen, B, Karlsen, AE, Kofod, H, Mølvig, J & Lernmark, A 1989, 'Site-specific antibodies distinguish single amino acid substitutions in position 57 in HLA-DQ beta-chain alleles associated with insulin-dependent diabetes', Journal of Immunology, vol. 143, no. 2, pp. 533-8.

APA

Atar, D., Dyrberg, T., Michelsen, B., Karlsen, A. E., Kofod, H., Mølvig, J., & Lernmark, A. (1989). Site-specific antibodies distinguish single amino acid substitutions in position 57 in HLA-DQ beta-chain alleles associated with insulin-dependent diabetes. Journal of Immunology, 143(2), 533-8.

Vancouver

Atar D, Dyrberg T, Michelsen B, Karlsen AE, Kofod H, Mølvig J et al. Site-specific antibodies distinguish single amino acid substitutions in position 57 in HLA-DQ beta-chain alleles associated with insulin-dependent diabetes. Journal of Immunology. 1989 Jul 15;143(2):533-8.

Author

Atar, D ; Dyrberg, T ; Michelsen, Birgitte ; Karlsen, Alan E ; Kofod, Hans ; Mølvig, J ; Lernmark, A. / Site-specific antibodies distinguish single amino acid substitutions in position 57 in HLA-DQ beta-chain alleles associated with insulin-dependent diabetes. In: Journal of Immunology. 1989 ; Vol. 143, No. 2. pp. 533-8.

Bibtex

@article{e25d623348b24edfb8b0637006383221,
title = "Site-specific antibodies distinguish single amino acid substitutions in position 57 in HLA-DQ beta-chain alleles associated with insulin-dependent diabetes",
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.",
keywords = "Alleles, Amino Acid Sequence, Antibody Specificity, Diabetes Mellitus, Type 1, Enzyme-Linked Immunosorbent Assay, HLA-DQ Antigens, Humans, Immune Sera, Immunoblotting, Molecular Sequence Data, Molecular Weight, Organ Specificity, Peptide Fragments",
author = "D Atar and T Dyrberg and Birgitte Michelsen and Karlsen, {Alan E} and Hans Kofod and J M{\o}lvig and A Lernmark",
year = "1989",
month = jul,
day = "15",
language = "English",
volume = "143",
pages = "533--8",
journal = "Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "2",

}

RIS

TY - JOUR

T1 - Site-specific antibodies distinguish single amino acid substitutions in position 57 in HLA-DQ beta-chain alleles associated with insulin-dependent diabetes

AU - Atar, D

AU - Dyrberg, T

AU - Michelsen, Birgitte

AU - Karlsen, Alan E

AU - Kofod, Hans

AU - Mølvig, J

AU - Lernmark, A

PY - 1989/7/15

Y1 - 1989/7/15

N2 - 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.

AB - 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.

KW - Alleles

KW - Amino Acid Sequence

KW - Antibody Specificity

KW - Diabetes Mellitus, Type 1

KW - Enzyme-Linked Immunosorbent Assay

KW - HLA-DQ Antigens

KW - Humans

KW - Immune Sera

KW - Immunoblotting

KW - Molecular Sequence Data

KW - Molecular Weight

KW - Organ Specificity

KW - Peptide Fragments

M3 - Journal article

C2 - 2738402

VL - 143

SP - 533

EP - 538

JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

IS - 2

ER -

ID: 45574977