Expression of the CTL2 transcript variants in human peripheral blood cells and human tissues
Expression of the CTL2 transcript variants in human peripheral blood cells and human tissues
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DOI：
10.1111/trf.12160
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发表时间：
2013-12-01
期刊：
影响因子：
2.9
通讯作者：
Bux, Juergen
中科院分区：
文献类型：
--
作者：
Flesch, Brigitte K.;Wesche, Jan;Bux, Juergen
研究背景HNA-3a抗原是输血相关急性肺损伤（TRALI）病理生理过程中的重要抗体靶点。它由胆碱转运蛋白样蛋白2（CTL 2）基因编码，该基因存在于两种转录变体TV 1和TV 2中，其上游启动子和编码区不同。只有TV 1已被证明，使胆碱运输穿过cellmembranes.Study设计和MethodsThe本研究的目的是确定在人类外周血细胞和组织中的CTL 2转录模式和它的能力结合HNA-3a抗体。从人全血、分离的中性粒细胞、单核血细胞、白细胞减少的血小板、人肺、肝和结肠中分离RNA。逆转录后，使用对相应转录物特异的引物组合扩增单链cDNA。含有转录变体TV 1或TV 2的整个CTL 2编码cDNA的质粒作为对照。HEK 293 T细胞表达这两种变体被用来确定HNA-3a antibodies.ResultsThe较短的TV 2转录证明在每个RNA样品来自人外周血测试到目前为止，以及在人肺和肝，而较长的TV 1转录只检测到在人肺和结肠。TV 1和TV 2具有相同的结合能力，HNA-3a antibodies. ConclusionTV 1和TV 2的表达是组织和细胞特异性的，与外周血细胞只表达TV 2。这不影响HNA-3a抗体的结合。这种不平等表达是否与TRALI的发病机制有关仍有待研究。
BackgroundThe HNA-3a antigen is an important antibody target in the pathophysiology of transfusion-related acute lung injury (TRALI). It is encoded by the choline transporter-like protein 2 (CTL2) gene, which exists in the two transcript variants TV1 and TV2, differing in the upstream promoter and coding region. Only TV1 has been demonstrated to enable choline transport across the cell membrane.Study Design and MethodsThe aim of this study was to determine the CTL2 transcript pattern in human peripheral blood cells and tissues and its capacity to bind HNA-3a antibodies. RNA was isolated from human whole blood, isolated neutrophils, mononuclear blood cells, leukoreduced platelets, human lung, liver, and colon. After reverse transcription, the single-stranded cDNA was amplified using primer combinations specific for the respective transcript. Plasmids containing the entire CTL2 coding cDNA of the transcript variant TV1 or TV2 served as controls. HEK293T cells expressing both variants were used to determine the binding of HNA-3a antibodies.ResultsThe shorter TV2 transcript was demonstrated in each RNA sample derived from human peripheral blood tested so far, as well as in human lung and liver, whereas the longer TV1 transcript was only detected in human lung and colon. TV1 and TV2 had the same binding capacity to HNA-3a antibodies.ConclusionThe expression of TV1 and TV2 is tissue and cell specific, with peripheral blood cells expressing only TV2. This does not affect binding of HNA-3a antibodies. Whether the unequal expression might be relevant in the pathogenesis of TRALI remains to be investigated.