Specific autoantigens identified by sera obtained from mice that are immunized with testicular germ cells alone.
Specific autoantigens identified by sera obtained from mice that are immunized with testicular germ cells alone.
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DOI：
10.1038/srep35599
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发表时间：
2016-10-18
影响因子：
4.6
通讯作者：
Itoh M
中科院分区：
文献类型：
--
作者：
Terayama H;Hirai S;Naito M;Qu N;Katagiri C;Nagahori K;Hayashi S;Sasaki H;Moriya S;Hiramoto M;Miyazawa K;Hatayama N;Li ZL;Sakabe K;Matsushita M;Itoh M
睾丸生殖细胞（TGCs）中存在多种自身免疫原性抗原（AIs），可被机体免疫系统识别为外来物。然而，关于tgc特异性AIs的信息很少。本研究的目的是鉴定tgc特异性AIs。我们之前已经证实，在不使用任何佐剂的情况下，使用活的同源TGC免疫也可以诱导小鼠实验性自身免疫性睾丸炎（EAO）。本研究通过TGC液相色谱-串联质谱分析鉴定TGC特异性AIs，并与EAO小鼠血清IgG进行双向凝胶电泳反应。在这项研究中，我们鉴定了11种tgc特异性AIs，它们与EAO小鼠的血清发生反应。Real-time RT-PCR分析显示，7种tgc特异性AIs的mRNA表达量仅在成熟睾丸中显著高于其他器官。此外，利用人胚胎肾293 （HEK293）细胞构建了10个tgc特异性AIs（未命名蛋白除外）的重组蛋白，并利用EAO血清反应进行Western blot验证。这些结果表明Atp6v1a、Hsc70t、Fbp1和Dazap1是tgc特异性AIs的候选物。这些ai的鉴定将为理解不孕症和癌症的发病机制提供新的途径，并可能为开发新的治疗方法提供基础。
There are various autoimmunogenic antigens (AIs) in testicular germ cells (TGCs) recognized as foreign by the body’s immune system. However, there is little information of TGC-specific AIs being available. The aim of this study is to identify TGC-specific AIs. We have previously established that immunization using viable syngeneic TGC can also induce murine experimental autoimmune orchitis (EAO) without using any adjuvant. This study is to identify TGC-specific AIs by TGC liquid chromatography–tandem mass spectrometry analysis, followed by two-dimensional gel electrophoresis that reacted with serum IgG from EAO mice. In this study, we identified 11 TGC-specific AIs that reacted with serum from EAO mice. Real-time RT-PCR analysis showed that the mRNA expressions of seven TGC-specific AIs were significantly higher in only mature testis compared to other organs. Moreover, the recombinant proteins of identified 10 (except unnamed protein) TGC-specific AIs were created by using human embryonic kidney 293 (HEK293) cells and these antigencities were reconfirmed by Western blot using EAO serum reaction. These results indicated Atp6v1a, Hsc70t, Fbp1 and Dazap1 were candidates for TGC-specific AIs. Identification of these AIs will facilitate new approaches for understanding infertility and cancer pathogenesis and may provide a basis for the development of novel therapies.