Boudreau Jeanette E, Le Luduec Jean-Benoît, Hsu Katharine C
Immunology Program, Sloan-Kettering Institute for Cancer Research, Memorial Sloan Kettering Cancer Center, New York, New York, United States of America.
Immunology Program, Sloan-Kettering Institute for Cancer Research, Memorial Sloan Kettering Cancer Center, New York, New York, United States of America; Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, New York, United States of America; Weill Medical College, Cornell University, New York, New York, United States of America.
PLoS One. 2014 Jun 11;9(6):e99543. doi: 10.1371/journal.pone.0099543. eCollection 2014.
Among NK cell receptor-ligand partnerships, KIR3DL1 and HLA-Bw4 demonstrate the greatest diversity; permutations of their allelic combinations titrate NK reactivity. Balancing selection has maintained distinct subtypes of KIR3DL1 alleles in global populations, implying that each may provide unique fitness advantages and variably influence disease processes. Though approaches exist for determining HLA-B allotypes, practical methods for identifying KIR3DL1 alleles are lacking. We have developed a PCR-based approach that identifies functional subtypes of KIR3DL1 alleles; it is suitable for research and may have clinical application. Six allele subsets were identified based on expression characteristics of the eleven most common KIR3DL1 alleles represented in reported populations. The remaining 62 low-frequency alleles were distributed into these groups based on sequence homology to coding regions. Subtype-specific SNPs were found in exons 3, 4, and 7, and used as priming sites for five multiplex PCR reactions. Genomic DNA derived from 175 unrelated donors and 52 related individuals from 6 families demonstrated >99.5% concordance between sequence-based typing and our novel approach. Finally, PCR-based typing accurately predicted NK phenotypes obtained by flow cytometry after staining with DX9 and Z27 monoclonal antibodies. This novel approach facilitates high-throughput analysis of KIR3DL1 allotypes to enable a broader understanding of KIR3DL1 and HLA-Bw4 interaction in health and disease.
在自然杀伤（NK）细胞受体 - 配体组合中，杀伤细胞免疫球蛋白样受体3DL1（KIR3DL1）和人类白细胞抗原Bw4（HLA - Bw4）表现出最大的多样性；它们等位基因组合的排列调节着NK细胞的反应性。平衡选择在全球人群中维持了KIR3DL1等位基因的不同亚型，这意味着每种亚型可能提供独特的适应性优势，并对疾病进程产生不同影响。虽然存在确定HLA - B亚型的方法，但缺乏识别KIR3DL1等位基因的实用方法。我们开发了一种基于聚合酶链反应（PCR）的方法来识别KIR3DL1等位基因的功能亚型；该方法适用于研究，可能也具有临床应用价值。根据已报道人群中11种最常见的KIR3DL1等位基因的表达特征，确定了6个等位基因子集。其余62个低频等位基因根据与编码区的序列同源性分配到这些组中。在第3、4和7外显子中发现了亚型特异性单核苷酸多态性（SNP），并将其用作5个多重PCR反应的引物位点。来自175名无关供体和6个家庭的52名相关个体的基因组DNA显示，基于序列的分型与我们的新方法之间的一致性>99.5%。最后，基于PCR的分型准确预测了用DX9和Z27单克隆抗体染色后通过流式细胞术获得的NK细胞表型。这种新方法有助于对KIR3DL1亚型进行高通量分析，从而更广泛地了解KIR3DL1和HLA - Bw4在健康和疾病中的相互作用。
