DNA typing for natural killer cell inhibiting HLA-Cw groups NK1 and NK2 by PCR-SSP
DNA typing for natural killer cell inhibiting HLA-Cw groups NK1 and NK2 by PCR-SSP
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DOI：
10.1016/s0022-1759(98)00126-4
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发表时间：
1998-09-01
影响因子：
2.2
通讯作者：
Kirchner, H
中科院分区：
文献类型：
--
作者：
Frohn, C;Schlenke, P;Kirchner, H
在过去的几年中，自然杀伤 (NK) 细胞已被证明可以表达 MHC 分子识别受体，这些受体被认为主要作为负信号受体发挥作用。 HLA-Cw 作为相应的配体似乎发挥着关键作用。氨基酸残基 77 和 80 不同的两个不同的 HLA-Cw 组抑制不同的 NK 细胞亚群。为了根据 HLA-Cw 组的表达对靶细胞进行分类，我们建立了组特异性 PCR-SSP，其直接扩增相关表位编码序列。通过重新分型从国际组织相容性研讨会获得的细胞系，并将这些结果与从我们肾移植单位的 80 对供体-受体对的等位基因特异性基因分型和血清分型获得的结果进行比较，对 PCR 方案进行了验证。在抑制性 HLA-Cw 受体的背景下，我们的方案明确区分了两个替代表位，是更直接、因此更可靠的方法，并且与等位基因特异性 PCR 或血清分型相比，劳动强度较小。此外，血清分型根本无法检测到某些等位基因。基础 NK 细胞研究和临床移植免疫学可能会受益于这种新建立的 PCR SSP 技术。 (C) 1998 Elsevier Science B.V. 保留所有权利。
Over the last few years, natural killer (NK) cells have been shown to express MHC molecule recognizing receptors which are thought to function primarily as negative signaling receptors. HLA-Cw seems to play a key role as the corresponding ligand. Two distinct HLA-Cw groups which differ in amino acid residues 77 and 80 inhibit separate subsets of NK cells. In order to classify target cells with respect to their expression of HLA-Cw groups we established a group specific PCR-SSP which directly amplifies the relevant epitope coding sequences. The PCR protocol was validated by retyping cell Lines obtained from the International Histocompatibility Workshop and by comparing those results with those acquired from allele-specific genotyping and serotyping on 80 donor-recipient pairs from our kidney transplantation unit. In the context of inhibitory HLA-Cw receptors, our protocol which definitively discriminates the two alternative epitopes is the more direct and thus more reliable approach, and is less labor intensive compared to an allele specific PCR or serotyping. In addition serotyping does not detect at all certain alleles. Basic NK cell research and clinical transplantation immunology may benefit from this newly established PCR SSP technique. (C) 1998 Elsevier Science B.V. All rights reserved.