Genomic identification of the minor histocompatibility antigen HA-1 locus by allele-specific PCR

M Wilke, J Pool, J M den Haan, E Goulmy

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Graft-versus-host disease (GvHD) can be a major complication of allogeneic bone marrow transplantation even in recipients of HLA genotype-identical transplants. Disparities in minor histocompatibility antigens (mHags) between donor and recipient are a potential risk for the development of GvHD. A mismatch for the mHag HA-1 can cause GvHD in adult recipients of allogeneic bone marrow from HLA-identical donors. The mHag HA-1, first identified by HLA-A*0201-restricted cytotoxic T cells (CTLs), was recently chemically characterized as a nonapeptide. On the cDNA level, the HA-1 locus has two alleles, HA-1H and HA-1R, which differ in two nucleotides, resulting in a single amino acid substitution. Here we report on the genomic structure of the HA-1 locus. Isolation and sequencing of cosmid DNA encoding the HA-1 peptide sequence revealed that the HA-1 alleles are encoded by two exons. Two different primer sets were designed, each consisting of allele-specific primers and a common primer, and both sets containing intronic sequences. We performed genomic DNA typing of three families consisting of 24 HLA-A*0201-positive individuals. The predicted allele-specific products correlated in all cases with the mHag classification by CTLs and by RT-PCR. We demonstrate for the first time the genomic identification of the mHag HA-1 locus. Prospective genomic typing for the HA-1 alleles will improve donor selection and identify HLA-A*0201-positive recipients with a high risk for HA-1-induced GvHD.

Original languageEnglish
Pages (from-to)312-7
Number of pages6
JournalTissue Antigens
Volume52
Issue number4
Publication statusPublished - Oct 1998

Cite this

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title = "Genomic identification of the minor histocompatibility antigen HA-1 locus by allele-specific PCR",
abstract = "Graft-versus-host disease (GvHD) can be a major complication of allogeneic bone marrow transplantation even in recipients of HLA genotype-identical transplants. Disparities in minor histocompatibility antigens (mHags) between donor and recipient are a potential risk for the development of GvHD. A mismatch for the mHag HA-1 can cause GvHD in adult recipients of allogeneic bone marrow from HLA-identical donors. The mHag HA-1, first identified by HLA-A*0201-restricted cytotoxic T cells (CTLs), was recently chemically characterized as a nonapeptide. On the cDNA level, the HA-1 locus has two alleles, HA-1H and HA-1R, which differ in two nucleotides, resulting in a single amino acid substitution. Here we report on the genomic structure of the HA-1 locus. Isolation and sequencing of cosmid DNA encoding the HA-1 peptide sequence revealed that the HA-1 alleles are encoded by two exons. Two different primer sets were designed, each consisting of allele-specific primers and a common primer, and both sets containing intronic sequences. We performed genomic DNA typing of three families consisting of 24 HLA-A*0201-positive individuals. The predicted allele-specific products correlated in all cases with the mHag classification by CTLs and by RT-PCR. We demonstrate for the first time the genomic identification of the mHag HA-1 locus. Prospective genomic typing for the HA-1 alleles will improve donor selection and identify HLA-A*0201-positive recipients with a high risk for HA-1-induced GvHD.",
keywords = "Adult, Alleles, Base Sequence, Cell Line, Chromosomes, Human, Pair 19, Cosmids/genetics, DNA Primers/metabolism, Graft vs Host Disease/genetics, Histocompatibility Testing, Humans, Minor Histocompatibility Antigens/genetics, Molecular Sequence Data, Oligopeptides/genetics, Polymerase Chain Reaction/methods",
author = "M Wilke and J Pool and {den Haan}, {J M} and E Goulmy",
year = "1998",
month = "10",
language = "English",
volume = "52",
pages = "312--7",
journal = "Tissue Antigens",
issn = "0001-2815",
publisher = "Wiley-Blackwell",
number = "4",

}

Genomic identification of the minor histocompatibility antigen HA-1 locus by allele-specific PCR. / Wilke, M; Pool, J; den Haan, J M; Goulmy, E.

In: Tissue Antigens, Vol. 52, No. 4, 10.1998, p. 312-7.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Genomic identification of the minor histocompatibility antigen HA-1 locus by allele-specific PCR

AU - Wilke, M

AU - Pool, J

AU - den Haan, J M

AU - Goulmy, E

PY - 1998/10

Y1 - 1998/10

N2 - Graft-versus-host disease (GvHD) can be a major complication of allogeneic bone marrow transplantation even in recipients of HLA genotype-identical transplants. Disparities in minor histocompatibility antigens (mHags) between donor and recipient are a potential risk for the development of GvHD. A mismatch for the mHag HA-1 can cause GvHD in adult recipients of allogeneic bone marrow from HLA-identical donors. The mHag HA-1, first identified by HLA-A*0201-restricted cytotoxic T cells (CTLs), was recently chemically characterized as a nonapeptide. On the cDNA level, the HA-1 locus has two alleles, HA-1H and HA-1R, which differ in two nucleotides, resulting in a single amino acid substitution. Here we report on the genomic structure of the HA-1 locus. Isolation and sequencing of cosmid DNA encoding the HA-1 peptide sequence revealed that the HA-1 alleles are encoded by two exons. Two different primer sets were designed, each consisting of allele-specific primers and a common primer, and both sets containing intronic sequences. We performed genomic DNA typing of three families consisting of 24 HLA-A*0201-positive individuals. The predicted allele-specific products correlated in all cases with the mHag classification by CTLs and by RT-PCR. We demonstrate for the first time the genomic identification of the mHag HA-1 locus. Prospective genomic typing for the HA-1 alleles will improve donor selection and identify HLA-A*0201-positive recipients with a high risk for HA-1-induced GvHD.

AB - Graft-versus-host disease (GvHD) can be a major complication of allogeneic bone marrow transplantation even in recipients of HLA genotype-identical transplants. Disparities in minor histocompatibility antigens (mHags) between donor and recipient are a potential risk for the development of GvHD. A mismatch for the mHag HA-1 can cause GvHD in adult recipients of allogeneic bone marrow from HLA-identical donors. The mHag HA-1, first identified by HLA-A*0201-restricted cytotoxic T cells (CTLs), was recently chemically characterized as a nonapeptide. On the cDNA level, the HA-1 locus has two alleles, HA-1H and HA-1R, which differ in two nucleotides, resulting in a single amino acid substitution. Here we report on the genomic structure of the HA-1 locus. Isolation and sequencing of cosmid DNA encoding the HA-1 peptide sequence revealed that the HA-1 alleles are encoded by two exons. Two different primer sets were designed, each consisting of allele-specific primers and a common primer, and both sets containing intronic sequences. We performed genomic DNA typing of three families consisting of 24 HLA-A*0201-positive individuals. The predicted allele-specific products correlated in all cases with the mHag classification by CTLs and by RT-PCR. We demonstrate for the first time the genomic identification of the mHag HA-1 locus. Prospective genomic typing for the HA-1 alleles will improve donor selection and identify HLA-A*0201-positive recipients with a high risk for HA-1-induced GvHD.

KW - Adult

KW - Alleles

KW - Base Sequence

KW - Cell Line

KW - Chromosomes, Human, Pair 19

KW - Cosmids/genetics

KW - DNA Primers/metabolism

KW - Graft vs Host Disease/genetics

KW - Histocompatibility Testing

KW - Humans

KW - Minor Histocompatibility Antigens/genetics

KW - Molecular Sequence Data

KW - Oligopeptides/genetics

KW - Polymerase Chain Reaction/methods

M3 - Article

VL - 52

SP - 312

EP - 317

JO - Tissue Antigens

JF - Tissue Antigens

SN - 0001-2815

IS - 4

ER -