Toll样受体及TAM受体酪氨酸激酶在调节小鼠睾丸免疫豁免中的功能

发布时间：2018-01-29 22:29

本文关键词： 睾丸免疫豁免自身免疫性睾丸炎 Toll样受体 TAM受体酪氨酸激酶　出处：《北京协和医学院》2015年博士论文　论文类型：学位论文

【摘要】：背景与目的：哺乳动物睾丸是一个典型的免疫豁免组织,系统性的免疫耐受和睾丸局部免疫抑制环境是维持免疫豁免的重要机制。生理状态下,生精细胞抗原在睾丸中不诱导免疫反应,但病理条件下,这种免疫平衡会被破坏,引起自身免疫性睾丸炎。睾丸局部免疫豁免及自身免疫性睾丸炎的调节机制有待深入研究。Toll样受体(TLR)在启动免疫反应中发挥重要作用,并可以介导自身免疫反应,而Tyro3, Axl与Mer (TAM)受体酪氨酸激酶能抑制TLR信号通路。本论文研究TLR与TAM对小鼠实验性自身免疫性睾丸炎(EAO)的调节作用,以揭示其在维持睾丸免疫豁免中的机制。材料方法：用完全佐剂乳化的睾丸匀浆免疫小鼠,诱导EAO模型。HE染色分析睾丸组织结构和损伤情况：免疫组化和流式细胞仪分析睾丸间质中巨噬细胞及淋巴细胞浸润情况；qRT-PCR和ELISA分析睾丸中促炎症因子表达；Western blot分析血清中自身抗体；生物素示踪的方法分析睾丸BTB结构完整性。结果：经过一次免疫同源睾丸组织匀浆,Axl和Mer双基因敲除(Ax1-/-Mer-/-)小鼠能诱导严重EAO。免疫50 d后,产生明显的EAO特征。淋巴结肿大,产生明显的系统性炎症反应；睾丸萎缩,重量减轻,生精上皮出现严重损伤：睾丸间质中出现明显的巨噬细胞和淋巴细胞浸润；睾丸内促炎症因子表达上调；血睾屏障被破坏；血清中产生抗生精细胞的自身抗体。经过一次诱导,Axl-/-和Mer-/-小鼠能产生相对弱的EAO,而在WT小鼠中不产生EAO。经过三次免疫,WT小鼠可产生严重EAO,出现典型的EAO表型。TLR2和TLR4单基因敲除(TLR2-/-和TLR4-/-)小鼠能诱导相对弱的EAO,而TLR2和TLR4双基因敲除(TLR2-/-TLR4-/-)小鼠不产生EAO。而且发现TLR2-/-和TLR2-/-TLR4-/-小鼠经过诱导后血清中没有产生自身抗体,表明TLR2在介导生殖细胞自身抗体产生中起关键作用。结论：TLR2和TLR4共同作用,介导EAO产生。而Axl和Mer协同作用,抑制EAO产生,这种抑制作用可能是通过抑制TLR2和TLR4信号通路实现的。研究结果进一步揭示了系统性免疫反应调节睾丸免疫豁免的机制。
[Abstract]:Background & objective: the mammalian testis are a typical immune immunity free tissue. Systemic immune tolerance and local immunosuppressive environment of testis are important mechanisms to maintain immunity immunity. Spermatogenic cell antigens do not induce immune responses in the testis, but this immune balance can be disrupted under pathological conditions. The local immunity immunity of testis and the regulatory mechanism of autoimmune orchitis need to be further studied. Toll-like receptor TLRR plays an important role in initiating the immune response. And can mediate autoimmune response while Tyro3. Axl and Mer tam receptor tyrosine kinase can inhibit TLR signaling pathway. In this study, we studied the regulatory effects of TLR and TAM on experimental autoimmune orchitis in mice. In order to reveal its mechanism in maintaining testicular immunity immunity. Material method: mice were immunized with complete adjuvant emulsified testis homogenate. EAO model. HE staining was used to analyze the structure and injury of testis: immunohistochemistry and flow cytometry were used to analyze the infiltration of macrophages and lymphocytes in testis stroma. QRT-PCR and ELISA were used to analyze the expression of proinflammatory factors in testis. Serum autoantibodies were analyzed by Western blot. The structural integrity of testicular BTB was analyzed by biotin tracer. Results: homologous homologous testis homogenate was immunized. Axl and Mer double gene knockout ax1-r-Mer-r-mice could induce severe EAO. After 50 days of immunization, they produced obvious EAO characteristics and enlarged lymph nodes. To produce obvious systemic inflammatory reaction; Testicular atrophy, weight reduction, spermatogenic epithelium appeared serious damage: clear interstitial macrophages and lymphocyte infiltration; The expression of proinflammatory factor was up-regulated in testis. The blood-testis barrier was destroyed; Autoantibodies against spermatogenic cells are produced in the serum. After one induction, axl-r-r-and Mer-r-mice produce relatively weak EAOs, whereas in WT mice, EAOs are not produced. WT mice can produce severe EAO, typical EAO phenotype. TLR2 and TLR4 single gene knockout TLR2-r- and TLR4-r-) mice can induce relatively weak EAO. And TLR2 and TLR4 double gene knockout TLR2-r-TLR4-r-). Mice did not produce EAO. and no autoantibodies were found in the serum of TLR2-r-r- and TLR2-r-TLR4-r- induced mice. It is suggested that TLR2 plays a key role in mediating the production of germ cell autoantibodies. Conclusion: TLR2 and TLR4 act together to mediate the production of EAO, while Axl and Mer play a synergistic role. Inhibition of EAO production may be achieved by inhibiting TLR2 and TLR4 signaling pathways. The results further reveal the mechanism of systemic immune response in regulating testicular immune immunity.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R392

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