The Type 1 Diabetes–Resistance Locus Idd22 Controls Trafficking of Autoreactive CTLs into the Pancreatic Islets of NOD Mice

Abstract

Type 1 Diabetes (T1D) has a strong genetic component. The Insulin dependent diabetes 22 (Idd22) locus was identified in crosses of T1D-susceptible NOD mice with the strongly T1D-resistant ALR strain. The NOD-Idd22 recombinant congenic mouse strain was generated where NOD mice carry the full Idd22 confidence interval. NOD-Idd22 mice exhibit almost complete protection from spontaneous T1D and a significant reduction in insulitis. Our goal was to unravel the mode of Idd22 based protection using in vivo and in vitro models. We determined that Idd22 did not impact immune cell diabetogenicity or β-cell resistance to cytoxicity in vitro. However, NOD-Idd22 mice were highly protected against adoptive transfer of T1D. Transferred CTL trafficked to the pancreatic lymph node (PLN) and proliferated to the same extent in NOD and NOD-Idd22 mice yet, the accumulation of pathogenic CTL in the islets was significantly reduced in NOD-Idd22 mice, correlating with disease resistance. Pancreatic endothelial cells from NOD-Idd22 animals expressed lower levels of adhesion molecules, even in response to inflammatory stimuli. Lower adhesion molecule expression resulted in weaker adherence of T cells to NOD-Idd22 endothelium as compared to NOD derived endothelium. Taken together, these results provide evidence that Idd22 regulates the ability of β-cell autoreactive T cells to traffic into the pancreatic islets and may represent a new target for pharmaceutical intervention to potentially prevent T1D.

Human CD8+T-cells Require Glycolysis to Elicit Effector Function

“…All mice used in this study were housed in specific pathogen-free facilities as previously described 20 . These studies were approved by the Institutional Animal Care and Use Committee (IACUC) at The Jackson Laboratories and University of Florida.…”

“…Flow cytometry analyses of the spleens of control and 2DG treated NOD/ShiLtJ mice, and peripheral blood and spleens of control and 2DG treated NOD.Ai4 mice, were carried out as previously described 20 . Blood samples were collected using EDTA coated tubes, and lymph nodes and spleens homogenized.…”

“…10-week-old female NOD/ShiLtJ were treated for 10 weeks with 35mM 2DG in their drinking water or placed on control water bottles prior to adoptive transfer. Splenocytes from NOD/ShiLtJ mice were isolated as previously described 20 . After isolation, splenocytes were pooled, washed, and resuspended in ice-cold Dulbecco’s PBS at a concentration of 100 × 10 6 /mL.…”

“…6-week-old NOD/ShiLtJ female mice were treated with 70mM 2DG in their drinking water or placed on control water bottles for 2 weeks prior to adoptive transfer. Splenocytes from NOD.Ai4 mice were isolated as previously described 20 and stained with CellTrace Violet (CTV) cell proliferation dye (ThermoFisher Scientific, USA), according to the manufacturer’s instructions. After labeling, splenocytes were washed and resuspended in ice-cold Dulbecco’s PBS at a concentration of 10 × 10 6 /mL.…”

NADPH Oxidase 2 Derived Reactive Oxygen Species Promote CD8+T cell Effector Function

“…T1D is caused by an autoimmune-mediated destruction of insulin-producing β cells (15)(16)(17)(18)(19)(20).…”