Wang W, Asiedu C, George J F, Thomas J M
Department of Surgery, University of Alabama at Birmingham 35294, USA.
Hum Immunol. 1998 Oct;59(10):599-606. doi: 10.1016/s0198-8859(98)00065-2.
The Fas/Fas ligand (FasL) interaction is pivotal in apoptosis-mediated regulation of the immune system. As such, it has relevance in areas of transplantation, gene therapy, AIDS, etc., all of which utilize the rhesus macaque as a preclinical animal model. In order to examine rhesus Fas/FasL, we cloned the rhesus FasL cDNA and have analyzed the function of the cloned gene. Our findings indicate that the rhesus FasL is highly homologous to the human but not the mouse (97% for human, 85% for mouse). In addition, soluble rhesus FasL can induce apoptosis, a property shared with the human soluble protein but not the mouse protein. The deduced protein sequence is 280 amino acids with a calculated Mr. of 31,646. Transfection of COS cells with the full-length cDNA yielded a 40 KD protein, which is in agreement with the size of human FasL. COS cells expressing rhesus FasL induced apoptosis in rhesus PHA blasts and human Fas+ CEM-6 cells. Thus, the cloned rhesus macaque FasL is functional and cross-reacts with human Fas. The cloned functional rhesus FasL cDNA will enable studies of its regulatory role in the nonhuman primate immune system.
Fas/Fas配体（FasL）相互作用在凋亡介导的免疫系统调节中起关键作用。因此，它在移植、基因治疗、艾滋病等领域具有相关性，所有这些领域都将恒河猴用作临床前动物模型。为了研究恒河猴Fas/FasL，我们克隆了恒河猴FasL cDNA并分析了克隆基因的功能。我们的研究结果表明，恒河猴FasL与人类高度同源，但与小鼠不同源（与人类同源性为97%，与小鼠同源性为85%）。此外，可溶性恒河猴FasL可诱导凋亡，这是人类可溶性蛋白所具有的特性，但小鼠蛋白不具有。推导的蛋白质序列为280个氨基酸，计算的分子量为31,646。用全长cDNA转染COS细胞产生了一种40 KD的蛋白质，这与人类FasL的大小一致。表达恒河猴FasL的COS细胞可诱导恒河猴PHA刺激的淋巴细胞和人类Fas+CEM-6细胞凋亡。因此，克隆的恒河猴FasL具有功能且与人Fas发生交叉反应。克隆的功能性恒河猴FasL cDNA将有助于研究其在非人灵长类动物免疫系统中的调节作用。
