Reddy S, Karanam M, Krissansen G, Nitschke K, Neve J, Poole C A, Ross J M
Department of Paediatrics, University of Auckland School of Medicine, New Zealand.
Histochem J. 2000 Apr;32(4):195-206. doi: 10.1023/a:1004084232446.
Beta cell destruction in NOD mice can be accelerated by adoptive transfer of diabetic spleen cells into irradiated adult NOD mice. Here mice receiving diabetic spleen cells were examined at days 0, 7, 14, 21 and at onset of diabetes for the resulting insulitis and the number of intra-islet CD4 and CD8 cells and macrophages. The progression of insulitis and the number of intra-islet CD4 and CD8 cells and macrophages were correlated with the expression and co-localization of inducible nitric oxide synthase, interferon-gamma and interleukin-4 by dual-label light and confocal immunofluorescence microscopy. Diabetes developed in 7/8 mice by 27 days following cell transfer. The insulitis score increased slightly by day 7 but rose sharply at day 14 (p = 0.001) and was maintained until diabetes. The mean number of intra-islet CD4 and CD8 cells and macrophages showed a similar trend to the insulitis scores and were present in almost equal numbers within the islets. Immunolabelling for inducible nitric oxide synthase was observed at day 7 in only some cells of a few islets but increased sharply from day 14. It was restricted to islets with insulitis and was co-localized in selective macrophages. Weak intra-islet interleukin-4 labelling was observed at days 7 and 14 but became more pronounced at day 21 and at onset of diabetes, being present in selective CD4 cells. Intra-islet labelling for interferon-gamma was first observed at day 21, but became more intense at onset of diabetes and was co-localized in a proportion of macrophages. Both cytokines were expressed in islets with advanced insulitis. Interferon-gamma staining was also observed within endothelial cells located in the exocrine pancreas. We conclude that transfer of diabetic spleen cells results in a rapid influx of CD4 and CD8 cells and macrophages within the pancreas of recipient mice. During the period of heightened insulitis, selective immune cells begin to express inducible nitric oxide synthase and the opposing cytokines, interferon-gamma and interleukin-4. Expression of these molecules becomes more pronounced immediately prior to and during the onset of diabetes.
将糖尿病小鼠的脾细胞过继转移到经辐照的成年非肥胖糖尿病（NOD）小鼠体内，可加速NOD小鼠的β细胞破坏。在此，对接受糖尿病小鼠脾细胞的小鼠在第0、7、14、21天以及糖尿病发病时进行检查，观察由此产生的胰岛炎以及胰岛内CD4和CD8细胞及巨噬细胞的数量。通过双标记光镜和共聚焦免疫荧光显微镜检查，将胰岛炎的进展以及胰岛内CD4和CD8细胞及巨噬细胞的数量与诱导型一氧化氮合酶、干扰素-γ和白细胞介素-4的表达及共定位进行关联分析。细胞转移后27天内，8只小鼠中有7只发生糖尿病。胰岛炎评分在第7天时略有增加，但在第14天急剧上升（p = 0.001），并一直维持到糖尿病发生。胰岛内CD4和CD8细胞及巨噬细胞的平均数量呈现出与胰岛炎评分相似的趋势，且在胰岛内的数量几乎相等。第7天时，仅在少数胰岛的部分细胞中观察到诱导型一氧化氮合酶的免疫标记，但从第14天起急剧增加。它局限于有胰岛炎的胰岛，并在选择性巨噬细胞中共定位。第7天和第14天时观察到胰岛内白细胞介素-4标记较弱，但在第21天和糖尿病发病时变得更加明显，存在于选择性CD4细胞中。胰岛内干扰素-γ标记最早在第21天观察到，但在糖尿病发病时变得更强烈，并在一部分巨噬细胞中共定位。两种细胞因子均在有晚期胰岛炎的胰岛中表达。在位于外分泌胰腺的内皮细胞中也观察到干扰素-γ染色。我们得出结论，糖尿病小鼠脾细胞的转移导致受体小鼠胰腺内CD4和CD8细胞及巨噬细胞迅速涌入。在胰岛炎加剧期间，选择性免疫细胞开始表达诱导型一氧化氮合酶以及相反的细胞因子，即干扰素-γ和白细胞介素-4。这些分子在糖尿病发病前及发病期间的表达变得更加明显。
