Dysregulated release and degradation of insulin during mononuclear cell-induced beta-cell lysis in HIT cells.
Activated human mononuclear cells (MCs) were coincubated for 8 h with HIT cells, a clonal cell line of pancreatic islet beta-cells. Measurements of HIT cell viability and insulin secretion were determined to 1) ascertain whether activated MCs can alter beta-cell viability in the absence of exogenously provided cytokines, 2) examine this response over a range of MC-HIT cell ratios, and 3) identify mechanisms responsible for altered insulin release consequent to MC-induced HIT cell damage. HIT cell viability was markedly decreased by activated MCs during an 8-h coincubation. HIT cell lysis could be attributed to activated natural killer cells, and lysis did not occur in the presence of activated T-lymphocyte clones. Activated MCs caused a marked early increase in insulin release from HIT cells (increase at 2 h: 7.75 +/- 0.16 nM for activated MCs, 2.66 +/- 0.09 nM for control; P less than 0.001). Insulin levels by the 8th h of the coincubation were significantly lower than the 2-h peak (4.33 +/- 0.13 vs. 7.75 +/- 0.16 nM, P less than 0.001). These changes in insulin were dependent on the ratio of activated MCs to HIT cells with the effects clearly evident at an activated MC-HIT cell ratio of greater than or equal to 10:1. Pretreatment of activated MCs and HIT cells with prostaglandin-synthesis inhibitors did not prevent the cytotoxic effects of activated MCs on HIT cells. Somatostatin did not inhibit the early exaggerated insulin release, suggesting that these increased insulin levels represented leakage of insulin from damaged HIT cells rather than functional insulin secretion.(ABSTRACT TRUNCATED AT 250 WORDS)
Robertson
R P
RP
Diabetes Center, UMHC, University of Minnesota, Minneapolis 55455.
Gromo
G
G
Zhang
H J
HJ
Walseth
T F
TF
Inverardi
L
L
D001095:Arachidonic Acids / Q000378:metabolism; D015704:CD4 Antigens / Q000032:analysis; D002450:Cell Communication; D002460:Cell Line; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007376:Interleukin-2 / Q000494:pharmacology; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D007694:Killer Cells, Natural / Q000502:physiology*; D007700:Kinetics; D007963:Leukocytes, Mononuclear / Q000502:physiology*; D008213:Lymphocyte Activation; D013004:Somatostatin / Q000494:pharmacology; D013601:T-Lymphocytes / Q000276:immunology / Q000502:physiology*
D001095:Arachidonic Acids; D015704:CD4 Antigens; D007328:Insulin; D007376:Interleukin-2; D013004:Somatostatin