title: Prevention of "Humanized" diabetogenic CD8 T-cell responses in HLA-transgenic NOD mice by a multipeptide coupled-cell approach.
abstract: OBJECTIVE
  Type 1 diabetes can be inhibited in standard NOD mice by autoantigen-specific immunotherapy targeting pathogenic CD8+ T-cells. NOD.β2m(null).HHD mice expressing human HLA-A2.1 but lacking murine major histocompatibility complex class I molecules develop diabetes characterized by CD8 T-cells recognizing certain autoantigenic peptides also targeted in human patients. These include peptides derived from the pancreatic β-cell proteins insulin (INS1/2 A(2-10) and INS1 B(5-14)) and islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP(265-273) and IGRP(228-236)). Hence, NOD.β2m(null).HHD mice represent a model system for developing potentially clinically translatable interventions for suppressing diabetogenic HLA-A2.1-restricted T-cell responses.
  
  
  METHODS
  Starting at 4-6 weeks of age, NOD.β2m(null).HHD female mice were injected intravenously with syngeneic splenocytes to which various admixtures of the four above-mentioned peptides were bound by the cross-linking agent ethylene carbodiimide (ECDI).
  
  
  RESULTS
  Treatment with such cells bearing the complete cocktail of INS and IGRP epitopes (designated INS/IGRP-SPs) significantly inhibited diabetes development in NOD.β2m(null).HHD recipients compared with controls receiving splenocytes coupled with an irrelevant HLA-A2.1-restricted Flu16 peptide. Subsequent analyses found syngeneic splenocytes bearing the combination of the two ECDI-coupled IGRPs but not INS peptides (IGRP-SPs or INS-SPs) effectively inhibited diabetes development in NOD.β2m(null).HHD mice. This result was supported by enzyme-linked immunospot (ELISPOT) analyses indicating combined INS/IGRP-SPs diminished HLA-A2.1-restricted IGRP but not INS autoreactive CD8+ T-cell responses in NOD.β2m(null).HHD mice.
  
  
  CONCLUSIONS
  These data support the potential of a cell therapy approach targeting HLA-A2.1-restricted IGRP autoreactive CD8 T-cells as a diabetes intervention approach in appropriate human patients.
authors.affiliation: Jackson Laboratory, Bar Harbor, Maine, USA.
authors.forename: Marijke
authors.initials: M
authors.lastname: Niens
authors.forename: Alexandra E
authors.initials: AE
authors.lastname: Grier
authors.forename: Michele
authors.initials: M
authors.lastname: Marron
authors.forename: Thomas W H
authors.initials: TW
authors.lastname: Kay
authors.forename: Dale L
authors.initials: DL
authors.lastname: Greiner
authors.forename: David V
authors.initials: DV
authors.lastname: Serreze
mesh_terms: D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002478:Cells, Cultured; D003922:Diabetes Mellitus, Type 1 / Q000378:metabolism / Q000628:therapy; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005952:Glucose-6-Phosphatase / Q000276:immunology; D015789:HLA-A2 Antigen / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007167:Immunotherapy; D051379:Mice; D016688:Mice, Inbred NOD; D008822:Mice, Transgenic; D011506:Proteins / Q000276:immunology; D013154:Spleen / Q000166:cytology
chemical_list: D015789:HLA-A2 Antigen; D011506:Proteins; D005952:Glucose-6-Phosphatase; C117983:G6pc2 protein, mouse