Antibodies to recoverin induce apoptosis of photoreceptor and bipolar cells in vivo
Antibodies to recoverin induce apoptosis of photoreceptor and bipolar cells in vivo
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DOI：
10.1006/jaut.1998.0221
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发表时间：
1998-10-01
影响因子：
12.8
通讯作者：
Fox, DA
中科院分区：
文献类型：
--
作者：
Adamus, G;Machnicki, M;Fox, DA
在癌症相关视网膜病变（CAR）综合征患者的血清中发现了针对恢复素的自身抗体。在这些研究中，我们检测了来自CAR患者血清的抗恢复素抗体和大鼠单克隆抗体对刘易斯大鼠视网膜的影响。玻璃体内注射后，抗恢复素自身抗体渗透到光感受器和双极细胞层中。它们在视网膜中的存在可以通过注射后24小时的免疫荧光检测到。与此同时，使用末端转移酶介导的dUTP缺口末端标记（TUNEL）和电子显微镜在整个光感受器和双极细胞层中识别出经历细胞凋亡的单个细胞。对照实验中使用的正常抗体不产生TUNEL标记。24 h时DNA梯状电泳证实DNA片段化。在电镜水平上，有明显的证据表明，细胞凋亡，细胞死亡的视网膜与抗恢复素抗体处理。在24和96小时，观察到核染色质凝聚和增加的空泡化的感光细胞外节。来自接受抗视网膜抗体的动物的视网膜的检查揭示了在外核层和内核层中1-2行的核的损失，而所有对照（假手术、正常IgG、磷酸盐缓冲盐水）显示了不变的核行数。此外，在用抗恢复蛋白抗体处理的视网膜中，外核层的核的行之间的间距增加，表明额外的细胞损失。这些研究提供了明确的证据表明，抗recoverin抗体能够穿透感光细胞和双极细胞，视网膜中recoverin表达的正常位点，并且抗recoverin抗体产生凋亡性细胞死亡。CAR患者也可能发生类似的机制，可能导致视力丧失和失明。(C)1998学术出版社
Autoantibodies against recoverin are found in the sera of patients with cancer-associated retinopathy (CAR) syndrome. In these studies we examined the effect of anti-recoverin antibodies from the sera of patients with CAR and rat monoclonal antibody on the retinas of Lewis rats. Anti-recoverin autoantibodies penetrated into the photoreceptor and bipolar cell layers following intravitreal injection. Their presence in the retina could be detected by immunofluorescence 24 h after injection. At the same time, individual cells undergoing apoptosis were identified throughout photoreceptor and bipolar cell layers using terminal transferase-mediated dUTP nick-end labeling (TUNEL) and electron microscopy. Normal antibodies used in control experiments did not produce TUNEL labeling. At 24 h, DNA fragmentation was confirmed by DNA ladder electrophoresis. At the electron microscopic level, there was clear evidence of cells undergoing apoptotic cell, death in the retinas treated with anti-recoverin antibodies,At 24 and 96 h, nuclear chromatin condensation and increased vacuolization of photoreceptor outer segments were observed. An examination of retinas from animals receiving anti-retinal antibodies revealed a loss of 1-2 rows of nuclei in the outer and inner nuclear layers whereas all controls (sham, normal IgG, phosphate buffered saline) showed an unchanged number of nuclei rows. In addition, there was an increase in spacing between the rows of nuclei of the outer nuclear layer in retinas treated with anti-recoverin antibodies, indicating additional cell loss. These studies provide clear evidence that anti-recoverin antibodies are capable of penetrating photoreceptor and bipolar cells, the normal site of recoverin expression in the retina, and that anti-recoverin antibodies produce apoptotic cell death. A similar mechanism may occur in patients with CAR, which may lead to visual loss and blindness. (C) 1998 Academic Press