Combined Genotypic and Phenotypic Killer Cell Ig-Like Receptor Analyses Reveal KIR2DL3 Alleles Displaying Unexpected Monoclonal Antibody Reactivity: Identification of the Amino Acid Residues Critical for Staining
Combined Genotypic and Phenotypic Killer Cell Ig-Like Receptor Analyses Reveal KIR2DL3 Alleles Displaying Unexpected Monoclonal Antibody Reactivity: Identification of the Amino Acid Residues Critical for Staining
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DOI：
10.4049/jimmunol.0903632
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发表时间：
2010-07-01
影响因子：
4.4
通讯作者：
Pende, Daniela
中科院分区：
文献类型：
--
作者：
Falco, Michela;Romeo, Elisa;Pende, Daniela
在人类，最近的临床和实验数据从造血干细胞移植显示，供体来源的同种异体反应性NK细胞发挥有益的移植物抗白血病的效果。根据供者杀伤细胞免疫球蛋白样受体（KIR）基因谱和供受者HLA I类分型，可以预测供者来源的同种异体反应性NK细胞的存在。此外，同种异体反应性NK细胞群体的大小可以通过组合使用抗KIR特异性mAb直接评估。在这项研究中，在试图提高同种异体反应性NK细胞亚群的定义，我们评估了KIR基因型和表型在一个队列的44名捐助者。该方法允许鉴定不与抗KIR 2DL 3特异性ECM 41 mAb反应的两种不同的KIR 2DL 3等位基因（KIR 2DL 3 *005和新等位基因KIR 2DL 3 *015）。相反，两个等位基因在细胞表面被与KIR 2DL 2/L3/S2反应的几种mAb识别。值得注意的是，KIR 2DL 3 *005也被抗KIR 2DL 1/S1特异性EB 6 B和11 PB 6 mAb染色。功能分析显示，尽管其特定的mAb反应性，KIR 2DL 3 *005对HLA-C分子的特异性与其他KIR 2DL 2/L3等位基因没有差异。最后，定点诱变证明，35位的谷氨酰胺是ECM 41染色所需的，而谷氨酸35和精氨酸50与EB 6 B或11 PB 6 mAb染色相关。我们目前的数据代表了一个实质性的进展，在表征的NK细胞库和一个改进的表型/功能定义给定的KIR+子集。免疫学杂志，2010，185：433-441。
In humans, recent clinical and experimental data from hematopoietic stem cell transplantation revealed that donor-derived alloreactive NK cells exert a beneficial graft versus leukemia effect. The existence of donor-derived alloreactive NK cells can be predicted on the basis of donor killer cell Ig-like receptor (KIR) gene profile and HLA class I typing of both donor and recipient. Moreover, the size of the alloreactive NK cell population can be directly assessed by the combined use of anti-KIR-specific mAb. In this study, in an attempt to improve the definition of alloreactive NK cell subsets, we assessed the KIR genotype and phenotype in a cohort of 44 donors. This approach allowed the identification of two different KIR2DL3 alleles (KIR2DL3*005 and the novel allele KIR2DL3*015) that did not react with the anti-KIR2DL3-specific ECM41 mAb. In contrast, both alleles were recognized at the cell surface by several mAb reacting with KIR2DL2/L3/S2. Notably, KIR2DL3*005 was also stained by the anti-KIR2DL1/S1-specific EB6B and 11PB6 mAb. Functional analysis revealed that, despite its particular mAb reactivity, the specificity of KIR2DL3*005 for HLA-C molecules did not differ from that of other KIR2DL2/L3 alleles. Finally, site-directed mutagenesis demonstrated that glutamine at position 35 is required for ECM41 staining, whereas glutamic acid 35 and arginine 50 are relevant for staining with EB6B or 11PB6 mAb. Our present data represent a substantial progress in the characterization of the NK cell repertoire and an improved phenotypic/functional definition of given KIR+ subsets. The Journal of Immunology, 2010, 185: 433-441.