Kniola B, Hellström S, Ludvigsson J
Department of Health and Environment, Linköping University Hospital, Sweden.
Diabetes Res Clin Pract. 1998 Mar;39(3):173-83. doi: 10.1016/s0168-8227(98)00009-6.
Islet cell surface antibodies (ICSA) have been demonstrated significantly more often in serum of patients with IDDM and their relatives than in healthy controls, but some investigators have been unable to show binding to human islets. It has therefore been suggested that ICSA is an artefact. We have localised and quantified ICSA binding to different specimens, including human pancreas. ICSA-positive and ICSA-negative patients' serum, determined by a RIA-method, were incubated with ultrathin sections of rat insulinoma (RIN5AH) cells, stained with Goat-anti-human IgG conjugated with 10 nm gold particles. Sections were viewed in transmission electron microscopy (Mag. 30,000 x) and image analyses was used to calculate immunolabelling. To test cell specificity we used (RIN5AH-cells, rat tumour cells producing growth hormone (GH3), normal human pancreas, human insulinoma, and mice liver). Sections showed good morphology. ICSA-positive sera, with or without islet cell antibodies (ICA) gave always higher immunocolloidal labelling then ICSA-negative sera on RIN5AH sections. Thus, the colloidal gold technique could confirm the ICSA results determined with RIA. Immunolabelling was pronounced on normal human pancreas and human insulinoma cells, but none was found on GH3-cells or mice liver cells. ICSA is not an artefact but do exist and bind to human islet cells and therefore maybe can be used as a marker for IDDM immunity.
与健康对照相比,胰岛细胞表面抗体(ICSA)在胰岛素依赖型糖尿病(IDDM)患者及其亲属血清中出现的频率明显更高,但一些研究人员未能证明其与人胰岛的结合。因此,有人提出ICSA是一种假象。我们已经对ICSA与包括人胰腺在内的不同标本的结合进行了定位和定量。通过放射免疫分析(RIA)方法确定的ICSA阳性和ICSA阴性患者血清,与用结合了10纳米金颗粒的山羊抗人IgG染色的大鼠胰岛素瘤(RIN5AH)细胞超薄切片一起孵育。切片在透射电子显微镜(放大倍数30,000倍)下观察,并使用图像分析来计算免疫标记。为了测试细胞特异性,我们使用了(RIN5AH细胞、产生生长激素的大鼠肿瘤细胞(GH3)、正常人胰腺、人胰岛素瘤和小鼠肝脏)。切片显示出良好的形态。在RIN5AH切片上,有或没有胰岛细胞抗体(ICA)的ICSA阳性血清的免疫胶体标记总是高于ICSA阴性血清。因此,胶体金技术可以证实用RIA测定的ICSA结果。在正常人胰腺和人胰岛素瘤细胞上免疫标记明显,但在GH3细胞或小鼠肝细胞上未发现。ICSA不是假象,确实存在并与人胰岛细胞结合,因此可能可用作IDDM免疫的标志物。