Identification of specific genes for untolerized testicular antigen in adult male mice

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  • 免疫寛容を受けていない精巣抗原遺伝子の同定

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<p> Spermatid and sperm do not appear in the seminiferous epithelium until puberty, when immune tolerance has already been established. Therefore, there are various autoimmunogenic antigens in testicular germ cells(TGC)recognized as foreign by the self-immune system. However, the blood-testis-barrier formed by Sertoli cells protect autoimmunogeneic spermatozoa from attack by the self-immune system. We previously showed that immunization of susceptible mice with TGC alone is sufficient to induce autoimmune orchitis without the use of any adjuvant. In the present study, we tried to identify TGC-specific autoantigens by phage display method, followed by mouse TGC phage library. The results showed that twenty genes were detected as candidate genes of autoantigens. Additionally, we analyzed more detailed gene expressions of these autoantigens by real-time PCR. Eight of twenty genes were specifically expressed in TGC. We generated plasmids with a Flag-tag of these nine candidate genes, and human embryonic kidney(HEK)293 T cells that transduced these plasmids were cultured to obtain these candidate gene expressing proteins. The proteins produced by HEK293T cells were reacted with TGC-immunized mouse serum antibody, and then detected by western blot analysis. As a result, two TGC-specific genes tested positive. These results indicate that these two TGC-specific genes are critical for induction of the autoimmune orchitis.</p>

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