Unit of Anatomy and Cell Biology, Department of Animal Sciences, Kyoto University, Kyoto, 606-8502, Japan,
Cytotechnology. 2000 Jul;33(1-3):189-201. doi: 10.1023/A:1008146119761.
Previously, we prepared an IgM monoclonal antibody(PFG-1) which specifically recognized a cell-membraneglycoprotein (PFG-1 antigen; 55 kD, pI 5.9),immunohistochemically reacted with granulosa cells ofhealthy follicles but not of atretic follicles, andinduced granulosa cell apoptosis. In the presentstudy, an IgM monoclonal antibody (PFG-3) capable ofinducing granulosa cell apoptosis and an IgGmonoclonal antibody (PFG-4) not capable of inducingapoptosis were produced against granulosa cellsprepared from healthy antral follicles of porcineovaries. Two-dimensional Western blotting analysisrevealed that PFG-3 specifically recognized twocell-membrane proteins (named PFG-3-1 andPFG-3-2/PFG-1 antigens; 42 kD, pI 5.2 and 55 kD, pI5.9, respectively) of healthy granulosa cells, andthat PFG-4 recognized the same two cell-membraneproteins. In atretic granulosa cells, PFG-3-2/PFG-1antigen disappeared. Immunochemical reactions of theseantibodies were only detected in follicular granulosacells but not any other ovarian tissues or organs.PFG-3 and PFG-4 immunohistochemically reacted withgranulosa cells of healthy and atretic follicles. Whenthe isolated granulosa cells prepared from healthyfollicles were cultured in medium containing PFG-3,the cells underwent apoptosis, and co-incubation withPFG-4 inhibited PFG-3-inducible apoptosis. Theseobservations suggested that PFG-3-2/PFG-1 antigen isa novel cell death receptor which is different fromthe apoptosis-mediating receptors (Fas/Apo-1/CD95 orTNF receptor), and that PFG-3-1 antigen may act as adecoy receptor and inhibit apoptotic signal transmission.
先前，我们制备了一种 IgM 单克隆抗体（PFG-1），它特异性地识别细胞膜糖蛋白（PFG-1 抗原；55kD，pI5.9），与健康卵泡的颗粒细胞发生免疫组织化学反应，但与闭锁卵泡的颗粒细胞无反应，并诱导颗粒细胞凋亡。在本研究中，针对来自猪卵巢健康窦状卵泡的颗粒细胞，制备了一种能够诱导颗粒细胞凋亡的 IgM 单克隆抗体（PFG-3）和一种不能诱导凋亡的 IgG 单克隆抗体（PFG-4）。二维 Western 印迹分析显示，PFG-3 特异性识别两种细胞膜蛋白（分别命名为 PFG-3-1 和 PFG-3-2/PFG-1 抗原；42kD，pI5.2 和 55kD，pI5.9），健康颗粒细胞，而 PFG-4 识别相同的两种细胞膜蛋白。在闭锁的颗粒细胞中，PFG-3-2/PFG-1 抗原消失。这些抗体的免疫化学反应仅在卵泡颗粒细胞中检测到，而在其他卵巢组织或器官中未检测到。PFG-3 和 PFG-4 免疫组织化学反应与健康和闭锁卵泡的颗粒细胞反应。当从健康卵泡中分离出的颗粒细胞在含有 PFG-3 的培养基中培养时，细胞发生凋亡，而与 PFG-4 共孵育可抑制 PFG-3 诱导的凋亡。这些观察结果表明，PFG-3-2/PFG-1 抗原是一种新型的细胞死亡受体，不同于凋亡介导的受体（Fas/Apo-1/CD95 或 TNF 受体），而 PFG-3-1 抗原可能作为诱饵受体并抑制凋亡信号的传递。
