Mari B, Breittmayer J P, Guerin S, Belhacene N, Peyron J F, Deckert M, Rossi B, Auberger P
INSERM U364, Faculté de Médecine, Nice, France.
Immunology. 1994 Jul;82(3):433-8.
Although it is now well established that cells of the immune system express most of the exopeptidases described so far, little information is available concerning the identification and the characterization of the peptidases associated with the surface of human thymocytes. In the present study we have focused on CD10 expression on thymocytes using both FACS and enzymatic analysis. Unfractionated intact human thymocytes were shown to express significant levels of CD10-specific enzymatic activity, as assessed by the hydrolysis of the neutral endopeptidase (NEP) substrate Suc-Ala-Ala-Phe-pNA and of D-Ala2-Leu-enkephalin, a typical NEP substrate. CD10 activity was abolished by specific NEP inhibitors, including thiorphan, retrothiorphan and phosphoramidon. Moreover, high performance liquid chromatography (HPLC) analysis showed that intact thymocytes and purified NEP hydrolysed thymopentin, a thymic factor known to induce the maturation of prothymocytes into thymocytes. Finally, CD 10/NEP was preferentially associated with CD3- CD3low and immature CD4- CD8- thymocytes. The data demonstrate for the first time that human thymocytes express functional NEP and suggest a role for this enzyme in the maturation of human thymocytes.
尽管目前已经明确免疫系统的细胞表达了迄今为止所描述的大多数外肽酶,但关于与人类胸腺细胞表面相关的肽酶的鉴定和表征的信息却很少。在本研究中,我们使用流式细胞术(FACS)和酶分析,重点研究了胸腺细胞上的CD10表达。通过中性内肽酶(NEP)底物Suc-Ala-Ala-Phe-pNA和典型的NEP底物D-Ala2-Leu-脑啡肽的水解评估,未分级的完整人类胸腺细胞显示出表达显著水平的CD10特异性酶活性。CD10活性被包括噻吗洛尔、逆转噻吗洛尔和磷酰胺在内的特异性NEP抑制剂所消除。此外,高效液相色谱(HPLC)分析表明,完整的胸腺细胞和纯化的NEP可水解胸腺五肽,胸腺五肽是一种已知可诱导前胸腺细胞成熟为胸腺细胞的胸腺因子。最后,CD10/NEP优先与CD3-CD3low和未成熟的CD4-CD8-胸腺细胞相关。这些数据首次证明人类胸腺细胞表达功能性NEP,并提示该酶在人类胸腺细胞成熟中起作用。