Faculty Research 1990 - 1999

NIT-1, a pancreatic beta-cell line established from a transgenic NOD/Lt mouse.

Document Type

Article

Publication Date

1991

Keywords

Animal, Antigens-Polyomavirus-Transforming: ge, Blotting-Northern, Cell-Line, Female, Glucagon: an, Glucose, Insulin: an, ge, se, Interferon-gamma-Recombinant, Islets-of-Langerhans: de, se, ul, Mice, Mice-Mutant-Strains, Mice-Transgenic, Microscopy-Electron, Pancreatic-Neoplasms, Promoter-Regions-(Genetics), Rats, RNA-Neoplasm: ge, SUPPORT-NON-U-S-GOVT, SUPPORT-U-S-GOVT-P-H-S, SV40-Virus: ge

First Page

842

Last Page

849

JAX Source

Diabetes 1991 Jul; 40(7):842-9.

Grant

DK36175, DK27722

Abstract

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 less than 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 Li 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.(ABSTRACT TRUNCATED AT 250 WORDS)

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