Rapid publication M. Wilke J. Pool J.M.M. den Haan E. Goulmy zyxwvutsr Key words: zyxwvutsrqponmlk bone marrow transplantation: genomic typing; graft-versus-host disease; HA-1; minor histocompatibility antigen Acknowledgments: We thank Drs zyxwvutsrqponm I? van den Elsen and G. M. Th. Schreuder for critical reading. We thank Dr. M. Giphart and A. Naipal for synthesis of the oligonucleotides. The human Cosmid Library was kindly provided by R. v. Haperen (Dept. of Cell Biology, EUR, Rotterdam, The Netherlands). Supported by the Dutch Cancer Foundation (KWF, to M. Wilke), the Dutch Organization for Scientific Research (NWO, to J.M.M. den Haan) and the J. A. Cohen Institute for Radiopathology and Radiation Protection (IRS). Received 17 August, accepted for publication 21 August 1 9 9 8 Copyright t3 Munlugaard 1998 zyxwvutsrqponm Wsue Antigens . ISSN 0001-2815 Tissue Antigens 1998: 52: 312-317 Printed in Denmark . All rights reserved Genomic identification of the minor histocompatibilityantigen HA-1 locus by allelespecific PCR Abstract: Graft-versus-host disease (GvHD) can be a major complication of allogeneic bone marrow transplantation even in recipients of HLA geno- type-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 recipi- ents 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-lHand HA-lR, which differ in two nucleo- tides, 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 zyxwvu HLA-A *0201-positive individ- uals. 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 gen- omic 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. Bone marrow transplantation (BMT) is the present treatment for hematological malignancies. One of the major drawbacks of allogen- eic BMT is graft-versus-host disease (GvHD). In recipients of allo- geneic HLA genotype-identical BMT, GvHD occurs in 15-35%, de- pending on the age of the recipient and the amount of T-cell de- pletion of the graft. The development of GvHD in recipients of HLA- identical bone martow (BiV) grafts reflects the role of minor histo- compatibility antigens (mHags) in BMT. Disparities for mHags be- tween HLA-identical BM donor and recipient pairs can evoke strong major histocompatibility complex (MHQrestricted cytotoxic and proliferative T-cell responses which can be measured zyxwv in zyxw vitm (re- Authors’ afflllatlon: M. Wllke. J. Pwl, J.M.M. den Haan. E. Goulmy Department of lmmunohernatologyand Bloodbank. Leiden University Medical Center (LUMC). Leiden. The Netherlands Correspondence to: Dr. M. Wilke Department of Bloodbank lmmunohematoiogy and Leiden University Medical Albinusdreef 2 2333 ZA Leiden The Netherlands TeI: 31-71.5263803 Fax: 31-71-5216751 E-mail: ihbsecr@euroneLnl Center 312