Effects of interleukin-12 in vitro on pancreatic islets isolated from normal rodents and from non-obese diabetic mice.
Administration of the T-helper 1 (Th 1) cell promoting cytokine interleukin-12 (IL-12) accelerates the development of autoimmune diabetes in non-obese diabetic (NOD) mice. In this study we examined the effects of IL-12 on isolated islets from NMRI (Naval Medical Research Institute-established) mice, Sprague-Dawley (S-D) rats and NOD mice. NMRI and S-D islets were cultured in medium RPMI 1640 + 10% fetal calf serum and exposed for 48 h to recombinant mouse IL-12 (0, 0.1, 1 and 10 ng/ml). Islet glucose metabolism, as measured by glucose oxidation rate, was suppressed by about 25% in NMRI islets exposed to 10 ng/ml IL-12. In rat islets 0.1 ng/ml IL-12 induced a 20% decrease in glucose oxidation rate. Islets cultured with 10 ng/ml IL-12 showed a decrease in medium insulin accumulation both in mouse and rat. Glucose-stimulated insulin release was lowered in rat islets exposed to 10 ng/ml IL-12, but not affected in NMRI islets. In NMRI islets IL-12 did not influence nitric oxide production as measured by nitrite formation. In rat islets IL-12 induced a decrease in nitrite formation compared with control islets. Islets were isolated from female NOD mice (age 5, 12, 20 and 26 weeks) and examined either immediately or cultured for 7 days with 10 ng/ml IL-12 alone or in combination with 4 ng/ml of the T-cell stimulating cytokine interleukin-2 (IL-2). In the age groups > 5 weeks of age the glucose-stimulated insulin release was lower in freshly isolated compared with cultured control islets. IL-2 + IL-12 addition induced a small decrease in glucose-stimulated insulin release in islets from 12-week-old animals. With increasing age the DNA content in freshly isolated islets increased due to immune cell infiltration. The DNA content in cultured islets was decreased by 40-60% compared with freshly isolated islets in the age groups over 5 weeks. Islet insulin content was similar in both freshly isolated and cultured islets. None of the cytokines, either alone or in combination, affected islet DNA or insulin content. We conclude that IL-12 has minor suppressive effects in vitro on normal rodent islets. It is likely that the reported accelerated diabetes development of IL-12 administration to NOD mice in vivo is not mediated by a direct toxic effect to the islets. The suppressed insulin release in NOD mouse islets treated with IL-2 + IL-12 suggests, however, that the accelerating effect might partly be attributed to stimulation of immune cells present in the insulitic lesion.
Department of Medical Cell Biology, Uppsala University Biomedicum, Sweden.
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J
Sternesjö
S
S
Sandler
D000375:Aging / Q000378:metabolism; D000818:Animals; D046508:Culture Techniques; D004247:DNA / Q000032:analysis; D003864:Depression, Chemical; D005260:Female; D005947:Glucose / Q000378:metabolism; D007328:Insulin / Q000032:analysis / Q000378:metabolism; D000078790:Insulin Secretion; D018664:Interleukin-12 / Q000494:pharmacology*; D007515:Islets of Langerhans / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D008815:Mice, Inbred Strains; D009573:Nitrites / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors
D007328:Insulin; D009573:Nitrites; D018664:Interleukin-12; D004247:DNA; D005947:Glucose