Full-length recombinant choline transporter-like protein 2 containing arginine 154 reconstitutes the epitope recognized by HNA-3a antibodies.
Full-length recombinant choline transporter-like protein 2 containing arginine 154 reconstitutes the epitope recognized by HNA-3a antibodies.
复制标题
含有精氨酸 154 的全长重组胆碱转运蛋白样蛋白 2 重建了 HNA-3a 抗体识别的表位。
DOI：
10.1111/j.1537-2995.2011.03411.x
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发表时间：
2012
期刊：
影响因子：
2.9
通讯作者：
Aster,RichardH
中科院分区：
文献类型：
--
作者：
Kanack,AdamJ;Peterson,JulieA;Sullivan,MiaJ;Bougie,DanielW;Curtis,BrianR;Aster,RichardH
背景：最近的报告显示，HNA-3a白细胞抗原（导致严重输血相关急性肺损伤的抗体的靶标）与胆碱转运蛋白样蛋白2（CTL 2）中的精氨酸154（而不是谷氨酰胺）多态性相关，但并未直接表明R154决定HNA-3a。迄今为止，含有R154的CTL 2肽仅被研究的一半HNA-3a抗体识别。研究设计和方法：HEK 293细胞用编码连接到绿色荧光蛋白（GFP）的全长CTL 2的cDNA转染。结果：20种HNA-3a抗体均优先与表达R154 CTL 2构建体的HEK 293细胞反应。HNA-3b抗体仅与CTL 2（Q154）反应。结论：这些发现提供了直接证据，证明在全长CTL 2的背景下，R154对于产生HNA-3a表位是必要且足够的，但表明蛋白质的翻译后修饰，例如，S-S键或添加聚糖，对于许多抗体识别HNA-3a是必要的。这可能会使大规模筛查献血者以检测抗HNA-3a的检测方法的开发复杂化。
BACKGROUND:Recent reports have shown that the HNA‐3a leukocyte antigen, a target for antibodies that cause severe transfusion‐related acute lung injury, correlates with an arginine 154 (rather than glutamine) polymorphism in choline transporter–like protein 2 (CTL2) but did not show directly that R154 determines HNA‐3a. CTL2 peptides containing R154 are recognized by only half of HNA‐3a antibodies studied to date. Constructs that react with all HNA‐3a antibodies are needed to fully define the HNA‐3a epitope.STUDY DESIGN AND METHODS:HEK293 cells were transfected with cDNA encoding full‐length CTL2 linked to green fluorescent protein (GFP). Transfectants were selected for GFP expression and tested with antibodies specific for HNA‐3a and ‐3b.RESULTS:Each of 20 HNA‐3a antibodies reacted preferentially with HEK293 cells expressing the R154 CTL2 construct. An HNA‐3b antibody reacted only with CTL2 (Q154).CONCLUSIONS:These findings provide direct evidence that R154 in the context of full‐length CTL2 is both necessary and sufficient to create the HNA‐3a epitope but suggest that posttranslational modifications of the protein, for example, S‐S bonds or addition of glycans, are necessary for recognition of HNA‐3a by many antibodies. This could complicate development of an assay for large‐scale screening of blood donors to detect anti‐HNA‐3a.