Willcox Abby, Gillespie Kathleen M
Diabetes and Metabolism Unit, School of Clinical Sciences, Southmead Hospital, University of Bristol, Bristol, UK.
Methods Mol Biol. 2016;1433:105-17. doi: 10.1007/7651_2015_287.
The islets of Langerhans play a critical role in glucose homeostasis. Islets are predominantly composed of insulin-secreting beta cells and glucagon-secreting alpha cells. In type 1 diabetes, the beta cells are destroyed by autoimmune destruction of insulin producing beta cells resulting in hyperglycemia. This is a gradual process, taking from several months to decades. Much of the beta cell destruction takes place during a silent, asymptomatic phase. Type 1 diabetes becomes clinically evident upon destruction of approximately 70-80 % of beta cell mass. Studying the decline in beta cell mass and the cells which are responsible for their demise is difficult as pancreatic biopsies are not feasible in patients with type 1 diabetes. The relative size of islets and their dispersed location throughout the pancreas means in vivo imaging of human islets is currently not manageable. At present, there are no validated biomarkers which accurately track the decline in beta cell mass in individuals who are at risk of developing, or have already developed, type 1 diabetes. Therefore, studies of pancreatic tissue retrieved at autopsy from donors with type 1 diabetes, or donors with high risk markers of type 1 diabetes such as circulating islet-associated autoantibodies, is currently the best method for studying beta cells and the associated inflammatory milieu in situ. In recent years, concerted efforts have been made to source such tissues for histological studies, enabling great insights to be made into the relationship between islets and the inflammatory insult to which they are subjected. This article describes in detail, a robust immunohistochemical method which can be utilized to study both recent, and archival human pancreatic tissue, in order to examine islet endocrine cells and the surrounding immune cells.
胰岛在葡萄糖稳态中发挥着关键作用。胰岛主要由分泌胰岛素的β细胞和分泌胰高血糖素的α细胞组成。在1型糖尿病中,胰岛素生成β细胞被自身免疫破坏,导致β细胞受损,进而引发高血糖。这是一个渐进的过程,从数月到数十年不等。大部分β细胞破坏发生在无症状的隐匿阶段。当大约70%-80%的β细胞团被破坏时,1型糖尿病才会在临床上显现出来。由于1型糖尿病患者无法进行胰腺活检,因此研究β细胞团的减少以及导致其死亡的细胞非常困难。胰岛的相对大小及其在整个胰腺中的分散位置意味着目前无法对人类胰岛进行体内成像。目前,尚无经过验证的生物标志物能够准确追踪有患1型糖尿病风险或已患1型糖尿病个体的β细胞团减少情况。因此,对1型糖尿病供体或具有1型糖尿病高风险标志物(如循环胰岛相关自身抗体)的供体尸检时获取的胰腺组织进行研究,是目前原位研究β细胞及相关炎症环境的最佳方法。近年来,人们齐心协力获取此类组织用于组织学研究,从而对胰岛与它们所遭受的炎症损伤之间的关系有了更深入的了解。本文详细描述了一种强大的免疫组织化学方法,可用于研究近期和存档的人类胰腺组织,以检查胰岛内分泌细胞和周围免疫细胞。