Release of glutamate decarboxylase-65 into the circulation by injured pancreatic islet beta-cells.
The enzyme glutamate decarboxylase-65 (GAD65) is a major autoantigen in autoimmune diabetes. The mechanism whereby autoreactivity to GAD65, an intracellular protein, is triggered is unknown, and it is possible that immunoreactive GAD65 is released by injured pancreatic islet beta-cells. There is a great need for methods by which to detect and monitor ongoing islet injury. If GAD65 were released and, furthermore, were able to reach the circulation, it could function as a marker of beta-cell injury. Here, a novel GAD65 plasma immunoassay is used to test the hypotheses that beta-cell injury induces GAD65 discharge in vivo and that discharged GAD65 reaches the bloodstream. Plasma GAD65 levels were determined in rats treated with alloxan, and with diabetogenic and low, subdiabetogenic doses of streptozotocin. beta-Cell injury resulted in GAD65 release into the circulation in a dose-dependent manner, and low-dose streptozotocin resulted in a more gradual increase in plasma GAD65 levels than did diabetogenic doses. Plasma GAD65 levels were reduced in rats that had undergone partial pancreatectomy and remained undetectable in mice. Together, these data demonstrate that GAD65 can be released into the circulation by injured beta-cells. Autoantigen shedding may contribute to the pathogenesis of islet autoimmunity in the multiple low-dose streptozocin model and perhaps, more generally, in other forms of autoimmune diabetes. These results demonstrate that, as is true with other tissues, islet injury, at least in some circumstances, can be monitored by use of discharged, circulating proteins. GAD65 is the first such confirmed protein marker of islet injury.
Waldrop
Megan A
MA
Department of Medicine, University of California-San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0726, USA.
Suckow
Arthur T
AT
Marcovina
Santica M
SM
Chessler
Steven D
SD
D000496:Alloxan / Q000008:administration & dosage / Q000494:pharmacology*; D000818:Animals; D015415:Biomarkers / Q000097:blood; D002096:C-Peptide / Q000097:blood; D016923:Cell Death; D004305:Dose-Response Relationship, Drug; D005968:Glutamate Decarboxylase / Q000097:blood*; D006207:Half-Life; D007274:Injections, Intraperitoneal; D007328:Insulin / Q000097:blood; D050417:Insulin-Secreting Cells / Q000187:drug effects* / Q000201:enzymology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010179:Pancreas / Q000187:drug effects / Q000473:pathology; D010180:Pancreatectomy; D051381:Rats; D017208:Rats, Wistar; D013311:Streptozocin / Q000008:administration & dosage / Q000494:pharmacology*
D015415:Biomarkers; D002096:C-Peptide; D007328:Insulin; D013311:Streptozocin; D000496:Alloxan; D005968:Glutamate Decarboxylase; C401141:glutamate decarboxylase 2