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

Authors

  • M. Wilke,

    Corresponding author
    1. Department of Immunohematology and Bloodbank, Leiden University Medical Center (LUMC), Leiden, The Netherlands
      Department of Immunohematology and Bloodbank Leiden University Medical Center Albinusdreef 2 2333 ZA Leiden The Netherlands Tel: 31–71-5263803 Fax: 31–71-5216751 E-mail: ihbsecr@euronet.nl
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  • J. Pool,

    1. Department of Immunohematology and Bloodbank, Leiden University Medical Center (LUMC), Leiden, The Netherlands
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  • J.M.M. den Haan,

    1. Department of Immunohematology and Bloodbank, Leiden University Medical Center (LUMC), Leiden, The Netherlands
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  • E. Goulmy

    1. Department of Immunohematology and Bloodbank, Leiden University Medical Center (LUMC), Leiden, The Netherlands
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Department of Immunohematology and Bloodbank Leiden University Medical Center Albinusdreef 2 2333 ZA Leiden The Netherlands Tel: 31–71-5263803 Fax: 31–71-5216751 E-mail: ihbsecr@euronet.nl

Abstract

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.

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