title: Macrophage contact dependent and independent TLR4 mechanisms induce β-cell dysfunction and apoptosis in a mouse model of type 2 diabetes.
abstract: Type 2 diabetes (T2D) is evolving into a global disease and patients have a systemic low-grade inflammation, yet the role of this inflammation is still not established. One plausible mechanism is enhanced expression and activity of the innate immune system. Therefore, we evaluated the expression and the function of the toll-like receptor 4 (TLR4) on pancreatic β-cells in primary mouse islets and on the murine β-cell line MIN6 in the presence or absence of macrophages. Diabetic islets have 40% fewer TLR4 positive β-cells, but twice the number of TLR4 positive macrophages as compared to healthy islets. Healthy and diabetic islets respond to a TLR4 challenge with enhanced production of cytokines (5-10-fold), while the TLR4 negative β-cell line MIN6 fails to produce cytokines. TLR4 stimulation induces β-cell dysfunction in mouse islets, measured as reduced glucose stimulated insulin secretion. Diabetic macrophages from 4-months old mice have acquired a transient enhanced capacity to produce cytokines when stimulated with LPS. Interestingly, this is lost in 6-months old diabetic mice. TLR4 activation alone does not induce apoptosis in islets or MIN-6 cells. In contrast, macrophages mediate TLR4-dependent cell-contact dependent (3-fold) as well as cell-contact independent (2-fold) apoptosis of both islets and MIN-6 cells. Importantly, diabetic macrophages have a significantly enhanced capacity to induce β-cell apoptosis compared to healthy macrophages. Taken together, the TLR4 responsiveness is elevated in the diabetic islets and mainly mediated by newly recruited macrophages. The TLR4 positive macrophages, in both a cell-contact dependent and independent manner, induce apoptosis of β-cells in a TLR4 dependent fashion and TLR4 activation directly induces β-cell dysfunction. Thus, targeting either the TLR4 pathway or the macrophages provides a novel attractive treatment regime for T2D.
authors.affiliation: Hagedorn Research Institute, Department of Diabetic Complication Biology, Måløv, Denmark.
authors.forename: Helena
authors.initials: H
authors.lastname: Cucak
authors.affiliation: Hagedorn Research Institute, Department of Diabetic Complication Biology, Måløv, Denmark.
authors.forename: Christopher
authors.initials: C
authors.lastname: Mayer
authors.affiliation: Hagedorn Research Institute, Department of Diabetic Complication Biology, Måløv, Denmark.
authors.forename: Morten
authors.initials: M
authors.lastname: Tonnesen
authors.affiliation: Hagedorn Research Institute, Department of Diabetic Complication Biology, Måløv, Denmark.
authors.forename: Lise Høj
authors.initials: LH
authors.lastname: Thomsen
authors.affiliation: Department of Incretin & Islet Biology, Måløv, Denmark.
authors.forename: Lars Groth
authors.initials: LG
authors.lastname: Grunnet
authors.affiliation: Hagedorn Research Institute, Department of Diabetic Complication Biology, Måløv, Denmark.
authors.forename: Alexander
authors.initials: A
authors.lastname: Rosendahl
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D016207:Cytokines / Q000378:metabolism; D003924:Diabetes Mellitus, Type 2 / Q000503:physiopathology*; D005260:Female; D005434:Flow Cytometry; D007118:Immunoassay; D007328:Insulin / Q000097:blood; D007515:Islets of Langerhans / Q000378:metabolism*; D008163:Luminescent Measurements; D008264:Macrophages / Q000378:metabolism*; D008297:Male; D051379:Mice; D051197:Toll-Like Receptor 4 / Q000276:immunology / Q000378:metabolism*
chemical_list: D016207:Cytokines; D007328:Insulin; C493487:Tlr4 protein, mouse; D051197:Toll-Like Receptor 4