Wilke M, Pool J, den Haan J M, Goulmy E
Department of Immunohematology and Bloodbank, Leiden University Medical Center, The Netherlands.
Tissue Antigens. 1998 Oct;52(4):312-7. doi: 10.1111/j.1399-0039.1998.tb03053.x.
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-A0201-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-A0201-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.
移植物抗宿主病（GvHD）可能是异基因骨髓移植的主要并发症，即使在接受HLA基因型相同移植的受者中也是如此。供体和受体之间次要组织相容性抗原（mHags）的差异是发生GvHD的潜在风险。mHag HA-1不匹配可导致来自HLA相同供体的异基因骨髓成年受者发生GvHD。mHag HA-1最初由HLA-A0201限制性细胞毒性T细胞（CTL）鉴定，最近经化学鉴定为一种九肽。在cDNA水平上，HA-1基因座有两个等位基因，HA-1H和HA-1R，它们在两个核苷酸上不同，导致单个氨基酸取代。在此我们报告HA-1基因座的基因组结构。对编码HA-1肽序列的黏粒DNA进行分离和测序，结果显示HA-1等位基因由两个外显子编码。设计了两组不同的引物，每组均由等位基因特异性引物和一个共同引物组成，且两组均包含内含子序列。我们对由24名HLA-A0201阳性个体组成的三个家族进行了基因组DNA分型。预测的等位基因特异性产物在所有情况下均与CTL和RT-PCR的mHag分类相关。我们首次证明了mHag HA-1基因座的基因组鉴定。对HA-1等位基因进行前瞻性基因组分型将改善供体选择，并识别出有高风险发生HA-1诱导的GvHD的HLA-A*0201阳性受者。
