Molecular Medicine, Istituto Giannina Gaslini, Genoa, Italy.
Hum Immunol. 2011 Oct;72(10):859-68. doi: 10.1016/j.humimm.2011.05.007. Epub 2011 May 24.
Specific combinations of killer immunoglobulin-like receptors (KIR) and human leukocyte antigen (HLA) class I molecules characterized by a particular residue 80 are significantly associated with outcomes in different pathologic conditions, such as autoimmunity, pathogenic infection, cancer, and reproductive failure. Thus, a simplified method for HLA typing used in association with the analysis of KIR genotype (Kirotype) is of particular interest to extend the analysis of larger series. Here, we describe a quick and inexpensive method that allows use of pyrosequencing, a helpful subtyping of HLA class I molecules, into HLA-Bw6, -Bw4 I(80) or -Bw4 T(80), HLA-C1, or -C2 groups and HLA-A allotypes sharing Bw4+ epitope or the rare HLA-B allotypes displaying the C1 motif. In particular, this analysis is focused on the amino acids around residue 80, known to be relevant in defining the affinity of KIR/HLA interaction and in the functional effects. This method was demonstrated to have good sensitivity, specificity, and reproducibility of detection and it was validated using a panel of HLA-typed International Histocompatibility Workshop (IHW) cell lines and clinical isolates. Using an allele quantitative acquisition mode, the method permitted us to obtain an accurate sequencing as required in heterozygous and/or homozygous sample definition.
特定的杀伤细胞免疫球蛋白样受体（KIR）和人类白细胞抗原（HLA）I 类分子组合，其特征是特定残基 80，与不同病理条件下的结果显著相关，如自身免疫、致病性感染、癌症和生殖失败。因此，一种与 KIR 基因型（Kirotype）分析相关的 HLA 分型简化方法特别有趣，可以扩展对更大系列的分析。在这里，我们描述了一种快速且廉价的方法，该方法允许使用焦磷酸测序，有助于 HLA I 类分子的亚分型，将 HLA-Bw6、-Bw4 I（80）或 -Bw4 T（80）、HLA-C1 或 -C2 组和 HLA-A 同种型共享 Bw4+表位或罕见的 HLA-B 同种型显示 C1 基序。特别是，该分析集中在已知与 KIR/HLA 相互作用亲和力和功能效应相关的残基 80 周围的氨基酸。该方法已被证明具有良好的灵敏度、特异性和可重复性，并且使用 HLA 分型的国际组织相容性工作会议（IHW）细胞系和临床分离株进行了验证。使用等位基因定量获取模式，该方法允许我们在杂合子和/或纯合子样本定义中获得所需的准确测序。
