NIT-1, A PANCREATIC BETA-CELL LINE ESTABLISHED FROM A TRANSGENIC NOD LT MOUSE
NIT-1, A PANCREATIC BETA-CELL LINE ESTABLISHED FROM A TRANSGENIC NOD LT MOUSE
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DOI：
10.2337/diabetes.40.7.842
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发表时间：
1991-07-01
期刊：
影响因子：
7.7
通讯作者：
LEITER, EH
中科院分区：
文献类型：
--
作者：
HAMAGUCHI, K;GASKINS, HR;LEITER, EH
携带大鼠胰岛素启动子/SV40大T抗原基因的NOD/Lt小鼠自发发生β细胞腺瘤。 NIT-1是从这些转基因小鼠之一建立的胰腺β细胞系。 第18代细胞免疫细胞化学染色显示大部分细胞含胰岛素，胰高血糖素含量<5%，不含胰多肽和生长抑素。 放免法测得细胞提取液中胰高血糖素含量仅为胰岛素含量的0.27%。 在16.5 mM葡萄糖下，两小时胰岛素分泌为638 ng/10（6）个细胞（细胞内含量的41%），而胰高血糖素仅为1.3 ng（细胞内含量的32%）。 在第11代和第19代之间，一致观察到对11和16.5 mM葡萄糖的响应刺激胰岛素分泌。 在第19代，茶碱和甲苯磺丁脲在5.5 mM葡萄糖刺激胰岛素分泌。 Northern印迹分析证实高水平的胰岛素mRNA，但只有微量胰高血糖素mRNA和不可检测的生长抑素mRNA。 干扰素-γ（IFN-γ）诱导的MHC I类RNA表达与细胞表面抗原表达显著增加相关。 类似地，仅在标准NOD/Lt胰岛细胞暴露于IFN-γ后通过Cr释放测定检测到的MHC-连接的"隐匿性" I类抗原在NIT-1细胞中被IFN-γ强烈诱导。 细胞表面MHC II类抗原在NIT-1细胞上没有组成型表达，并且在IFN-γ孵育后无法检测到，尽管证明了IFN-γ诱导的Aa、Ab和Ii不变链RNA转录本。 类似地，IFN-γ诱导细胞间粘附分子1（Icam-1）转录物并不伴有ICAM-1抗原的可证实的细胞表面表达。 NIT-1细胞表现出分化小鼠β细胞的典型超微结构特征，包括许多β颗粒。 一个不寻常的发现是NIT-1细胞从其表面脱落成熟的C型逆转录病毒。 由于该细胞系代表了永生化NOD β细胞基因组的来源，因此它将用于处理β细胞抗原的各种研究。
NOD/Lt mice harboring a hybrid rat insulin-promoter/SV40 large T-antigen gene spontaneously develop beta-cell adenomas. NIT-1 is a pancreatic beta-cell line established from one of these transgenic mice. Immunocytochemical staining of passage 18 cells showed most contained insulin, with < 5% containing glucagon, and none containing pancreatic polypeptide or somatostatin. Glucagon content radioimmunoassayed in cell extracts was only 0.27% of the insulin content. Two-hour insulin secretion at 16.5 mM glucose was 638 ng/10(6) cells (41% of intracellular content) compared to only 1.3 ng glucagon (32% of intracellular content). Stimulated insulin secretion was consistently observed in response to 11 and 16.5 mM glucose between passages 11 and 19. At passage 19, both theophylline and tolbutamide stimulated insulin secretion at 5.5 mM glucose. Northern-blot analysis confirmed high levels of insulin mRNA but only trace glucagon mRNA and undetectable somatostatin mRNA. Interferon-gamma (IFN-gamma)-induced MHC class I RNA expression was correlated with markedly increased antigen expression at the cell surface. Similarly, a MHC-linked "occult" class I-like antigen detected by Cr release assay only after exposure of standard NOD/Lt islet cells to IFN-gamma was strongly induced by IFN-gamma in NIT-1 cells. Cell surface MHC class II antigen was not constitutively expressed on NIT-1 cells and could not be detected after IFN-gamma incubation, despite demonstration of IFN-gamma-induced Aa, Ab, and Ii invariant-chain RNA transcripts. Similarly IFN-gamma induction of intercellular adhesion molecule 1 (Icam-1) transcripts was not accompanied by demonstrable cell surface expression of ICAM-1 antigen. NIT-1 cells exhibited ultrastructural features typical of differentiated mouse beta-cells, including numerous beta-granules. An unusual finding was that NIT-1 cells were shedding mature type C retrovirus from their surfaces. Because this cell line represents a source of immortalized NOD beta-cell genome, it will be useful for a wide variety of studies dealing with beta-cell antigens.