In vitro inhibition of insulin release by blood mononuclear cells from insulin-dependent diabetic and healthy subjects: synergistic action of IL-1 and TNF.
Previous studies have demonstrated that peripheral blood mononuclear cells (BMC) from type 1 (insulin-dependent) diabetic patients inhibit insulin release (IR) from rat or mouse islet cells in vitro. This phenomenon is of great interest as a model for islet graft rejection. We found that lipopolysaccharide (LPS)-stimulated BMC of healthy donors and type 1 diabetic patients suppress both basal and stimulated insulin secretion. To study whether this inhibition was due to soluble mediators we added supernatants of LPS-stimulated BMC or recombinant human interleukin-1 beta (IL-1) and tumor necrosis factor-alpha (TNF) at concentrations comparable to those found in the supernatants to rat islet cells. The inhibitory effect of BMC on islet cells could be transferred by supernatants of LPS-stimulated BMC. We found that neither IL-1 nor TNF alone inhibit IR from dispersed adult rat islet cells. However, the combination of IL-1 and TNF was highly effective. Ultrafiltration of supernatants of LPS-stimulated BMC through a PM-10 membrane (10 kDa cutoff) deprived the supernatants of the inhibitory activity indicating that only intact IL-1 and TNF (m.w. about 17 kDa), but not smaller IL-1 and TNF fragments, were responsible for the effects on islet cells. These data suggest that activation of BMC and cytokine release at islet graft site may result in an early loss of graft function. Islet transplantation using microcapsules not permeable for molecules with m.w. > 10 kDa would be preferable.
Ablamunits
V
V
Institute of Transplantology and Artificial Organs, Laboratory of Immunology, Moscow, Russia.
Baranova
F
F
Mandrup-Poulsen
T
T
Nerup
J
J
D000328:Adult; D000818:Animals; D002478:Cells, Cultured; D003922:Diabetes Mellitus, Type 1 / Q000097:blood*; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D004926:Escherichia coli; D005260:Female; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007375:Interleukin-1 / Q000494:pharmacology*; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000633:toxicity; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000494:pharmacology; D012016:Reference Values; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D007328:Insulin; D007375:Interleukin-1; D008070:Lipopolysaccharides; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha