A comprehensive analysis of the binding of anti-MR antibodies to activating KIRs
A comprehensive analysis of the binding of anti-MR antibodies to activating KIRs
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DOI：
10.1038/gene.2013.58
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发表时间：
2014-01-01
影响因子：
5
通讯作者：
Gonzalez, A.
中科院分区：
文献类型：
--
作者：
Czaja, K.;Borer, A-S;Gonzalez, A.
由于可用抗体的交叉反应性，特别是激活性和抑制性同工型之间的交叉反应性，杀伤细胞免疫球蛋白样受体 (KIR) 表达的分析一直非常困难。我们对与激活 KIR (a-KIR) 结合的可用抗 KIR 抗体进行了全面研究。使用稳定转染 a-KIR（KIR2DS1-S5 和 KIR3DS1）的细胞系，我们证实了记录的结合特异性。此外，我们还发现克隆 HPMA4 和 143211（之前被认为对 KIR2DS1/L1 和 KIR2DL1 具有特异性）分别结合 KIR2DS5 和 KIR2DS3 (HPMA4) 以及 KIR2DS5 (143211)。其他具有先前未记录的结合的抗体是 JJC11.6（识别 KIR2DS3）和 5.133（识别除 KIR2DS1 和 KIR2DS3 之外的所有 a-KIR）。通过用可溶性 KIR-Fc 融合蛋白封闭以及对分选的原代自然杀伤细胞进行逆转录酶 PCR 分析，证实了新的 KIR2DS5 反应性。总之，我们展示了以前未记录的抗 KIR 抗体的结合特性。分析 KIR 表达时应考虑这些交叉反应。
Analysis of killer cell immunoglobulin-like receptor (KIR) expression has been notoriously difficult because of the cross-reactivity of available antibodies, in particular between activating and inhibitory isoforms. We undertook a comprehensive study of available anti-KIR antibodies binding to activating KIRs (a-KIRs). Using cell lines stably transfected with a-KIRs (KIR2DS1-S5 and KIR3DS1), we confirmed documented binding specificities. In addition, we show that clones HPMA4 and 143211-previously assumed to be specific for KIR2DS1/L1 and KIR2DL1, respectively-bind KIR2DS5 and KIR2DS3 (HPMA4), and KIR2DS5 (143211). Other antibodies with previously undocumented binding were JJC11.6 (recognizing KIR2DS3) and 5.133 (recognizing all a-KIRs except KIR2DS1 and KIR2DS3). The novel KIR2DS5 reactivities were confirmed by blocking with soluble KIR-Fc fusion proteins, and by reverse transcriptase-PCR analysis of sorted primary natural killer cells. In conclusion, we show formerly undocumented binding properties of anti-KIR antibodies. These cross-reactivities should be taken into account when analyzing KIR expression.