Rajalingam Raja, Ashouri Elham
Department of Pathology and Laboratory Medicine, UCLA Immunogenetics Center, David Geffen School of Medicine at UCLA, University of California at Los Angeles, Los Angeles, CA, USA.
Methods Mol Biol. 2013;1034:239-55. doi: 10.1007/978-1-62703-493-7_12.
By interacting with specific HLA class I molecules, the killer cell immunoglobulin-like receptors (KIR) regulate the effector function of natural killer (NK) cells and subsets of CD8 T cells. The KIR receptors and HLA class I ligands are encoded by unlinked polymorphic gene families located on different human chromosomes, 19 and 6, respectively. The number and type of KIR genes are substantially variable between individuals, which may contribute to human diversity in responding to infection, malignancy and allogeneic transplants. PCR typing using sequence-specific primers (PCR-SSP) is the most commonly used method to determine KIR gene content. This chapter describes a step-by-step protocol for PCR-SSP typing to identify the presence and absence of all 16 known KIR genes. Moreover, the chapter provides the basic rules to verify the accuracy of KIR genotyping results and explains specific methods for the data analysis.
通过与特定的I类人类白细胞抗原(HLA)分子相互作用,杀伤细胞免疫球蛋白样受体(KIR)调节自然杀伤(NK)细胞和CD8 T细胞亚群的效应功能。KIR受体和I类HLA配体分别由位于不同人类染色体(19号和6号)上的不连锁多态基因家族编码。个体之间KIR基因的数量和类型存在很大差异,这可能导致人类在应对感染、恶性肿瘤和同种异体移植方面的多样性。使用序列特异性引物的聚合酶链反应(PCR-SSP)分型是确定KIR基因组成最常用的方法。本章描述了用于PCR-SSP分型以鉴定所有16个已知KIR基因存在与否的逐步方案。此外,本章提供了验证KIR基因分型结果准确性的基本规则,并解释了数据分析的具体方法。