Cell surface trafficking of Fas in NIT-1 cells and dissection of surface and total Fas expression.
The appearance of Fas receptor at the surface of pancreatic beta-cells affected by progressive insulitis strongly suggests that Fas-mediated beta-cell apoptosis plays an important role in the pathogenesis of type 1 diabetes. In support of this concept, the present study has shown that islet cells from NOD mice and the beta-cell line NIT-1 respond to the proinflammatory cytokines IL-1beta and IFN-gamma with Fas surface expression in a dose- and time-dependent manner. Moreover, the prevention of cytokine-induced surface Fas expression by actinomycin D, cycloheximide, and brefeldin A demonstrated that trafficking of Fas to the beta-cell surface requires RNA and protein synthesis and, in addition is critically dependent on intracellular protein transport. Compared with total cellular Fas protein, the amount of Fas at the cell surface was relatively small and indicated that Fas is preferentially expressed in cytoplasmic compartments of NIT-1 cells. It is concluded that inflammatory insults specifically induce translocation of Fas to the beta-cell surface and that interference with cell surface Fas expression is a new strategy to improve beta-cell survival in inflamed islets.
Augstein
P
P
Gerhardt Katsch Institute of Diabetes-Karlsburg e. V., Greifswalder Strasse 11e, 17495 Karlsburg, Germany. augstein@mail.uni-greifswald.de
Dunger
A
A
Salzsieder
C
C
Heinke
P
P
Kubernath
R
R
Bahr
J
J
Fischer
U
U
Rettig
R
R
Salzsieder
E
E
D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D020126:Brefeldin A / Q000494:pharmacology; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D018920:Coculture Techniques; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D004171:Diploidy; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D007249:Inflammation; D007371:Interferon-gamma / Q000378:metabolism; D007375:Interleukin-1 / Q000378:metabolism; D007515:Islets of Langerhans / Q000378:metabolism*; D051379:Mice; D016688:Mice, Inbred NOD; D009569:Nitric Oxide / Q000378:metabolism; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D014176:Protein Biosynthesis; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D021381:Protein Transport; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D013997:Time Factors; D014407:Tumor Cells, Cultured; D019014:fas Receptor / Q000096:biosynthesis*
D007375:Interleukin-1; D019384:Nucleic Acid Synthesis Inhibitors; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D019014:fas Receptor; D003609:Dactinomycin; D020126:Brefeldin A; D009569:Nitric Oxide; D012313:RNA; D007371:Interferon-gamma; D003513:Cycloheximide