Lebedeva Tatiana V, Ohashi Marina, Zannelli Georgia, Cullen Rebecca, Yu Neng
HLA Laboratory, American Red Cross Blood Services, Northeast Division, Dedham, Massachusetts, USA.
Hum Immunol. 2007 Sep;68(9):789-96. doi: 10.1016/j.humimm.2007.07.002. Epub 2007 Aug 1.
Multiple studies suggest that prospective KIR typing may be beneficial for the outcome of bone marrow transplants, but to date no practical high-resolution KIR typing system has been developed. Here we propose a comprehensive high-resolution typing approach that provides allele level KIR typing. Based on the low-resolution typing obtained by SSO, the 14 KIR loci are divided in groups according to the level of polymorphism in exons coding for extracellular Ig-like domains and cytoplasmic tail. The first group is typed by sequence-specific oligonucleotide only; the second is typed by sequence-based typing (SBT) based on the amplification of a fragment coding the Ig-like domains; and the third is typed by SBT based on amplification of a fragment coding the cytoplasmic tail. SBT for the fourth group includes both the Ig-like and cytoplasmic domains. Because of a considerable number of polymorphisms scattered throughout all nine KIR exons, SBT results may still produce a number of ambiguities, which can be resolved by sequence-specific primers. This combined high-resolution approach was applied to the complete KIR typing of 205 Caucasian hematopoietic stem cell donors in support of the National Marrow Donor Program High-resolution KIR Typing Pilot Project. High-resolution typing of several KIR loci produced numerous novel alleles, whereas some loci demonstrated very limited polymorphism. Several of the novel alleles appeared in more than four donors, suggesting that these alleles are not rare. Our results showed that the comprehensive KIR typing approach presented here provides the balance of high-resolution typing and cost effectiveness.
多项研究表明，前瞻性KIR分型可能对骨髓移植的结果有益，但迄今为止尚未开发出实用的高分辨率KIR分型系统。在此，我们提出了一种全面的高分辨率分型方法，可提供等位基因水平的KIR分型。基于通过序列特异性寡核苷酸（SSO）获得的低分辨率分型，根据编码细胞外免疫球蛋白样结构域和胞质尾的外显子中的多态性水平，将14个KIR基因座分为几组。第一组仅通过序列特异性寡核苷酸进行分型；第二组通过基于编码免疫球蛋白样结构域的片段扩增的序列分型（SBT）进行分型；第三组通过基于编码胞质尾的片段扩增的SBT进行分型。第四组的SBT包括免疫球蛋白样结构域和胞质结构域。由于在所有九个KIR外显子中散布着大量多态性，SBT结果可能仍会产生一些模糊性，这可以通过序列特异性引物来解决。这种组合的高分辨率方法被应用于205名白种人造血干细胞供体的完整KIR分型，以支持国家骨髓捐献计划高分辨率KIR分型试点项目。几个KIR基因座的高分辨率分型产生了许多新的等位基因，而一些基因座显示出非常有限的多态性。其中几个新等位基因出现在四名以上的供体中，表明这些等位基因并不罕见。我们的结果表明，本文提出的全面KIR分型方法在高分辨率分型和成本效益之间取得了平衡。
