The Type 1 Diabetes PhysioLab® Platform: a validated physiologically based mathematical model of pathogenesis in the non‐obese diabetic mouse

Type 1 diabetes is an autoimmune disease whose clinical onset signifies a lifelong requirement for insulin therapy and increased risk of medical complications. To increase the efficiency and confidence with which drug candidates advance to human type 1 diabetes clinical trials, we have generated and validated a mathematical model of type 1 diabetes pathophysiology in a well‐characterized animal model of spontaneous type 1 diabetes, the non‐obese diabetic (NOD) mouse. The model is based on an extensive survey of the public literature and input from an independent scientific advisory board. It reproduces key disease features including activation and expansion of autoreactive lymphocytes in the pancreatic lymph nodes (PLNs), islet infiltration and β cell loss leading to hyperglycaemia. The model uses ordinary differential and algebraic equations to represent the pancreas and PLN as well as dynamic interactions of multiple cell types (e.g. dendritic cells, macrophages, CD4+ T lymphocytes, CD8+ T lymphocytes, regulatory T cells, β cells). The simulated features of untreated pathogenesis and disease outcomes for multiple interventions compare favourably with published experimental data. Thus, a mathematical model reproducing type 1 diabetes pathophysiology in the NOD mouse, validated based on accurate reproduction of results from multiple published interventions, is available for in silico hypothesis testing. Predictive biosimulation research evaluating therapeutic strategies and underlying biological mechanisms is intended to deprioritize hypotheses that impact disease outcome weakly and focus experimental research on hypotheses likely to provide insight into the disease and its treatment.

[1]  C. Ferran,et al.  DEPLETING ANTI-CD4 MONOCLONAL ANTIBODY CURES NEW-ONSET DIABETES, PREVENTS RECURRENT AUTOIMMUNE DIABETES, AND DELAYS ALLOGRAFT REJECTION IN NONOBESE DIABETIC MICE1 , 2004, Transplantation.

[2]  R. Flavell,et al.  Helper Requirements for Generation of Effector CTL to Islet β Cell Antigens1 , 2004, The Journal of Immunology.

[3]  Polly Matzinger,et al.  'Educated' dendritic cells act as messengers from memory to naive T helper cells , 2004, Nature Immunology.

[4]  J. Rosmalen,et al.  Sex Steroids Influence Pancreatic Islet Hypertrophy and Subsequent Autoimmune Infiltration in Nonobese Diabetic (NOD) and NODscid Mice , 2001, Laboratory Investigation.

[5]  M. Nakayama,et al.  Tolerance mechanisms in murine autoimmune diabetes induced by anti-ICAM-1/LFA-1 mAb and anti-CD8 mAb. , 2002, The Kobe journal of medical sciences.

[6]  J. Bluestone,et al.  Impairment of NK cell function by NKG2D modulation in NOD mice. , 2003, Immunity.

[7]  Y. Hamamoto,et al.  Guidelines for computer modeling of diabetes and its complications. , 2004, Diabetes care.

[8]  G. Eisenbarth,et al.  Prime role for an insulin epitope in the development of type 1 diabetes in NOD mice , 2005, Nature.

[9]  D. Umetsu,et al.  Pulmonary dendritic cells producing IL-10 mediate tolerance induced by respiratory exposure to antigen , 2001, Nature Immunology.

[10]  Yuehong Wu,et al.  Increased Nonobese Diabetic Th1:Th2 (IFN-γ:IL-4) Ratio Is CD4+ T Cell Intrinsic and Independent of APC Genetic Background1 , 2002, The Journal of Immunology.

[11]  F. Wong,et al.  Pancreatic infiltration but not diabetes occurs in the relative absence of MHC class II-restricted CD4 T cells: studies using NOD/CIITA-deficient mice. , 1999, Journal of immunology.

[12]  W. Wheat,et al.  Increased NF‐κB activity in B cells and bone marrow‐derived dendritic cells from NOD mice , 2004 .

[13]  M. Kasuga,et al.  Suppression and acceleration of autoimmune diabetes by neutralization of endogenous interleukin-12 in NOD mice. , 2000, Diabetes.

[14]  D. Godfrey,et al.  Flow cytometric study of T cell development in NOD mice reveals a deficiency in alphabetaTCR+CDR-CD8- thymocytes. , 1997, Journal of autoimmunity.

[15]  B. Rocha,et al.  Peripheral Selection of  T Cell Repertoires: The Role of Continuous Thymus Output , 1997, The Journal of experimental medicine.

[16]  David Gray,et al.  B cells regulate autoimmunity by provision of IL-10 , 2002, Nature Immunology.

[17]  K. Polonsky,et al.  Increased beta-cell proliferation and reduced mass before diabetes onset in the nonobese diabetic mouse. , 1999, Diabetes.

[18]  Damien Bresson,et al.  Sa.65. Biosimulations Predict Optimal Oral Insulin/Anti-CD3 and Oral Insulin/Exendin-4 Combination Treatment Regimens for the Reversal of Diabetes in the Non-Obese Diabetic (NOD) Mouse , 2008 .

[19]  C. Piccirillo,et al.  TGF-β1 Somatic Gene Therapy Prevents Autoimmune Disease in Nonobese Diabetic Mice , 1998, The Journal of Immunology.

[20]  Z. Dai,et al.  Cutting Edge: Secondary Lymphoid Organs Are Essential for Maintaining the CD4, But Not CD8, Naive T Cell Pool1 , 2001, The Journal of Immunology.

[21]  Jeffrey A. Bluestone,et al.  In Vitro–expanded Antigen-specific Regulatory T Cells Suppress Autoimmune Diabetes , 2004, The Journal of experimental medicine.

[22]  P. Ricciardi-Castagnoli,et al.  Dendritic cells directly trigger NK cell functions: Cross-talk relevant in innate anti-tumor immune responses in vivo , 1999, Nature Medicine.

[23]  W. Winter,et al.  Flow cytometric enumeration of mononuclear cell populations infiltrating the islets of Langerhans in prediabetic NOD mice: development of a model of autoimmune insulitis for type I diabetes. , 1990, Regional immunology.

[24]  J. Habener,et al.  Cure of overt diabetes in NOD mice by transient treatment with anti-lymphocyte serum and exendin-4. , 2004, Diabetes.

[25]  A. Pombo,et al.  Dendritic cells interact directly with naive B lymphocytes to transfer antigen and initiate class switching in a primary T-dependent response. , 1998, Journal of immunology.

[26]  P. Santamaria.  Kinetic Evolution of a Diabetogenic CD8+ T Cell Response , 2003, Annals of the New York Academy of Sciences.

[27]  B. Woda,et al.  NOD mice have a generalized defect in their response to transplantation tolerance induction. , 1999, Diabetes.

[28]  D. Coleman,et al.  The non-obese diabetic (NOD) mouse. , 1987, The American journal of pathology.

[29]  D. Finegood,et al.  Phagocytosis of apoptotic cells by macrophages from NOD mice is reduced. , 2002, Diabetes.

[30]  A. M. Shapiro,et al.  Combination therapy with low dose sirolimus and tacrolimus is synergistic in preventing spontaneous and recurrent autoimmune diabetes in non-obese diabetic mice , 2002, Diabetologia.

[31]  Y. Z. Ider,et al.  Quantitative estimation of insulin sensitivity. , 1979, The American journal of physiology.

[32]  R. Flavell,et al.  Helper requirements for generation of effector CTL to islet beta cell antigens. , 2004, Journal of immunology.

[33]  P. Brunetti,et al.  Pancreatic beta-cell destruction in non-obese diabetic mice. , 1993, Metabolism: clinical and experimental.

[34]  Ping Zhang,et al.  Guidelines for computer modeling of diabetes and its complications , 2004 .

[35]  C. Benoist,et al.  The role of CD8+ T cells in the initiation of insulin‐dependent diabetes mellitus , 1996, European journal of immunology.

[36]  J. Novak,et al.  NKT cells inhibit the onset of diabetes by impairing the development of pathogenic T cells specific for pancreatic beta cells. , 2002, Immunity.

[37]  Saroja Ramanujan,et al.  A comprehensive review of interventions in the NOD mouse and implications for translation. , 2005, Immunity.

[38]  H. Kolb,et al.  The APC1 concept of type I diabetes. , 1998, Autoimmunity.

[39]  C. Benoist,et al.  Initiation of Autoimmune Diabetes by Developmentally Regulated Presentation of Islet Cell Antigens in the Pancreatic Lymph Nodes , 1999, The Journal of experimental medicine.

[40]  R. Tisch,et al.  L-Selectin Is Not Required for T Cell-Mediated Autoimmune Diabetes1 , 2002, The Journal of Immunology.

[41]  Y. Matsuzawa,et al.  The NOD mouse. , 1994, Diabetes research and clinical practice.

[42]  J. Bluestone,et al.  The role of CD28 and CTLA4 in the function and homeostasis of CD4+CD25+ regulatory T cells. , 2003, Novartis Foundation symposium.

[43]  R. Tisch,et al.  CD40 ligand-CD40 interactions are necessary for the initiation of insulitis and diabetes in nonobese diabetic mice. , 1997, Journal of immunology.

[44]  M. Toublanc,et al.  Course of pancreatic beta cell destruction in prediabetic NOD mice: a histomorphometric evaluation. , 1994, Diabete & metabolisme.

[45]  C. Benoist,et al.  Physiological β Cell Death Triggers Priming of Self-reactive T Cells by Dendritic Cells in a Type-1 Diabetes Model , 2003, The Journal of experimental medicine.

[46]  L. Karlsson,et al.  Evaluation of phagocytic activity in human monocyte-derived dendritic cells. , 2001, Journal of aerosol medicine : the official journal of the International Society for Aerosols in Medicine.

[47]  Shimon Sakaguchi,et al.  Homeostatic maintenance of natural Foxp3 + CD25+ CD4+ regulatory T cells by interleukin (IL)-2 and induction of autoimmune disease by IL-2 neutralization , 2005, The Journal of experimental medicine.

[48]  R. Schreiber,et al.  IFN-gamma action on pancreatic beta cells causes class I MHC upregulation but not diabetes. , 1998, The Journal of clinical investigation.

[49]  J. Levine,et al.  Cytokine Dysregulation Induced by Apoptotic Cells Is a Shared Characteristic of Macrophages from Nonobese Diabetic and Systemic Lupus Erythematosus-Prone Mice 1 , 2004, The Journal of Immunology.

[50]  D. Godfrey,et al.  Flow cytometric study of T cell development in NOD mice reveals a deficiency in αβTCR+CD4-CD8- thymocytes. , 1997 .

[51]  G. Szot,et al.  Costimulation controls diabetes by altering the balance of pathogenic and regulatory T cells. , 2004, The Journal of clinical investigation.

[52]  P. Ricciardi-Castagnoli,et al.  Fcγ Receptor–mediated Induction of Dendritic Cell Maturation and Major Histocompatibility Complex Class I–restricted Antigen Presentation after Immune Complex Internalization , 1999, The Journal of experimental medicine.

[53]  W. Wheat,et al.  Increased NF-kappa B activity in B cells and bone marrow-derived dendritic cells from NOD mice. , 2004, European journal of immunology.

[54]  L. Chatenoud,et al.  [Remission of established disease in diabetic NOD mice induced by anti-CD3 monoclonal antibody]. , 1992, Comptes rendus de l'Academie des sciences. Serie III, Sciences de la vie.

[55]  B. Pakkenberg,et al.  Genetic background determines the size and structure of the endocrine pancreas. , 2005, Diabetes.

[56]  L. Moretta,et al.  NK-dependent DC maturation is mediated by TNFalpha and IFNgamma released upon engagement of the NKp30 triggering receptor. , 2005, Blood.

[57]  T. Saito,et al.  Interferon gamma production by natural killer (NK) cells and NK1.1+ T cells upon NKR-P1 cross-linking , 1996, The Journal of experimental medicine.

[58]  J. Gudjonsson,et al.  The Role of CD8 T Cells and Their Antigen Receptors in Psoriasis , 2010 .

[59]  L. Leserman,et al.  Class I‐restricted presentation of exogenous antigen acquired by Fcγ receptor‐mediated endocytosis is regulated in dendritic cells , 2000, European journal of immunology.

[60]  M. Rewers,et al.  Early expression of antiinsulin autoantibodies of humans and the NOD mouse: evidence for early determination of subsequent diabetes. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[61]  Christopher R. Myers,et al.  Universally Sloppy Parameter Sensitivities in Systems Biology Models , 2007, PLoS Comput. Biol..

[62]  D. Wagner,et al.  Peripheral CD4loCD40+ auto‐aggressive T cell expansion during insulin‐dependent diabetes mellitus , 2004, European journal of immunology.

[63]  C. Ricordi,et al.  Prolonged islet graft survival in NOD mice by blockade of the CD40-CD154 pathway of T-cell costimulation. , 2001, Diabetes.

[64]  J. Allison,et al.  CD28-mediated costimulation is necessary for optimal proliferation of murine NK cells. , 1994, Journal of immunology.

[65]  C. Benoist,et al.  T-cell compartments of prediabetic NOD mice. , 2003, Diabetes.

[66]  R. Kastelein,et al.  Comparison of the effects of interleukin-1 alpha, interleukin-1 beta and interferon-gamma-inducing factor on the production of interferon-gamma by natural killer. , 1997, European journal of immunology.

[67]  Mark S. Anderson,et al.  The NOD mouse: a model of immune dysregulation. , 2005, Annual review of immunology.

[68]  A. Signore,et al.  The natural history of lymphocyte subsets infiltrating the pancreas of NOD mice , 1989, Diabetologia.

[69]  J. Sredy,et al.  Rapamycin prevents the onset of insulin‐dependent diabetes mellitus (IDDM) in NOD mice , 1992, Clinical and experimental immunology.

[70]  R. Tisch,et al.  Immune response to glutamic acid decarboxylase correlates with insulitis in non-obese diabetic mice , 1993, Nature.

[71]  N. Shastri,et al.  Ligands for the murine NKG2D receptor: expression by tumor cells and activation of NK cells and macrophages , 2000, Nature Immunology.

[72]  P. Brunetti,et al.  Pancreatic β-cell destruction in non-obese diabetic mice , 1993 .

[73]  L. Chatenoud,et al.  Anti-CD3 antibody induces long-term remission of overt autoimmunity in nonobese diabetic mice. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[74]  F. Sutterwala,et al.  Transforming growth factor-beta controls T helper type 1 cell development through regulation of natural killer cell interferon-gamma. , 2005, Nature immunology.

[75]  L. Adorini,et al.  Dynamics of Pathogenic and Suppressor T Cells in Autoimmune Diabetes Development , 2003, The Journal of Immunology.

[76]  J. Bluestone,et al.  Differential effects of anti-B7-1 and anti-B7-2 monoclonal antibody treatment on the development of diabetes in the nonobese diabetic mouse , 1995, The Journal of experimental medicine.

[77]  H. Drexhage,et al.  Dendritic cells and macrophages are essential for the retention of lymphocytes in (peri)-insulitis of the nonobese diabetic mouse: a phagocyte depletion study , 2005, Laboratory Investigation.

[78]  M. Monahan,et al.  Recombinant human IL-10 prevents the onset of diabetes in the nonobese diabetic mouse. , 1994, Clinical immunology and immunopathology.

[79]  R. Kastelein,et al.  Comparison of the effects of interleukin‐1α, interleukin‐lβ and interferon‐γ‐inducing factor on the production of interferon‐γ by natural killer , 1997 .

[80]  D. Allan,et al.  Apoptosis Is the Mode of β-Cell Death Responsible for the Development of IDDM in the Nonobese Diabetic (NOD) Mouse , 1997, Diabetes.

[81]  A. Naji,et al.  Impaired Activation of Islet-Reactive CD4 T Cells in Pancreatic Lymph Nodes of B Cell-Deficient Nonobese Diabetic Mice1 , 2001, The Journal of Immunology.

[82]  R. Tisch,et al.  Elevated NF-κB Activation in Nonobese Diabetic Mouse Dendritic Cells Results in Enhanced APC Function1 , 2002, The Journal of Immunology.

[83]  P. Morel,et al.  Phenotypic and functional characteristics of BM-derived DC from NOD and non-diabetes-prone strains. , 2001, Clinical immunology.

[84]  David M Eddy,et al.  Archimedes: a trial-validated model of diabetes. , 2003, Diabetes care.

[85]  F. Lepault,et al.  Expression of homing and adhesion molecules in infiltrated islets of Langerhans and salivary glands of nonobese diabetic mice. , 1994, Journal of immunology.

[86]  J. Corbett,et al.  Interleukin-1 Plus γ-Interferon-Induced Pancreatic β-Cell Dysfunction Is Mediated by β-Cell Nitric Oxide Production , 2002 .

[87]  G. Hedlund,et al.  Dendritic cells and macrophages are the first and major producers of TNF-alpha in pancreatic islets in the nonobese diabetic mouse. , 1998, Journal of immunology.

[88]  A. Hayward,et al.  Reduced incidence of insulitis in NOD mice following anti-CD3 injection: requirement for neonatal injection. , 1992, Journal of autoimmunity.

[89]  J. Rosmalen,et al.  Abnormalities in dendritic cell and macrophage accumulation in the pancreas of nonobese diabetic (NOD) mice during the early neonatal period. , 2002, Histology and histopathology.

[90]  L. Edelstein-Keshet,et al.  Quantifying macrophage defects in type 1 diabetes. , 2005, Journal of theoretical biology.

[91]  H. Ljunggren,et al.  Triggering of murine NK cells by CD40 and CD86 (B7-2). , 1999, Journal of immunology.

[92]  C. Fathman,et al.  Immunotherapy of the nonobese diabetic mouse: treatment with an antibody to T-helper lymphocytes. , 1988, Science.

[93]  D. Becker,et al.  Autoimmune islet destruction in spontaneous type 1 diabetes is not β-cell exclusive , 2003, Nature Medicine.

[94]  M. Smyth,et al.  Cytometric and functional analyses of NK and NKT cell deficiencies in NOD mice. , 2001, International immunology.

[95]  K. Gadkar,et al.  Mechanisms Mediating Anti‐CD3 Antibody Efficacy , 2005, Annals of the New York Academy of Sciences.

[96]  A. Shapiro,et al.  Costimulation blockade of both inducible costimulator and CD40 ligand induces dominant tolerance to islet allografts and prevents spontaneous autoimmune diabetes in the NOD mouse. , 2006, Diabetes.

[97]  D. Finegood,et al.  Clearance of apoptotic β-cells is reduced in neonatal autoimmune diabetes-prone rats , 2002, Cell Death and Differentiation.

[98]  S. Bonner-Weir,et al.  Dynamics of β-cell Mass in the Growing Rat Pancreas: Estimation With a Simple Mathematical Model , 1995, Diabetes.

[99]  H. Hall,et al.  Broadly impaired NK cell function in non-obese diabetic mice is partially restored by NK cell activation in vivo and by IL-12/IL-18 in vitro. , 2004, International immunology.

[100]  K. Rock,et al.  Efficient major histocompatibility complex class I presentation of exogenous antigen upon phagocytosis by macrophages. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[101]  P. Snow,et al.  Induction of Autoantigen-Specific Th2 and Tr1 Regulatory T Cells and Modulation of Autoimmune Diabetes1 , 2003, The Journal of Immunology.

[102]  D. Greiner,et al.  Multiple defects in innate and adaptive immunologic function in NOD/LtSz-scid mice. , 1995, Journal of immunology.

[103]  B. Göke,et al.  Glucagon-like-peptide-1 (7–36) amide improves glucose sensitivity in beta-cells of NOD mice , 1996, Acta Diabetologica.

[104]  M. Kasuga,et al.  Administration of IL-4 prevents autoimmune diabetes but enhances pancreatic insulitis in NOD mice. , 1998, Clinical immunology and immunopathology.

[105]  T. Utsugi,et al.  Perforin-independent beta-cell destruction by diabetogenic CD8(+) T lymphocytes in transgenic nonobese diabetic mice. , 1999, The Journal of clinical investigation.

[106]  D. Finegood,et al.  Neonatal beta-cell apoptosis: a trigger for autoimmune diabetes? , 2000, Diabetes.

[107]  H. Jun,et al.  The Role of Macrophages in T Cell–mediated Autoimmune Diabetes in Nonobese Diabetic Mice , 1999, The Journal of experimental medicine.

[108]  J. Sprent,et al.  Life span of naive and memory t cells , 1995, Stem cells.

[109]  Saroja Ramanujan,et al.  Dosing and Timing Effects of Anti‐CD40L Therapy , 2007, Annals of the New York Academy of Sciences.

[110]  P. Mottram,et al.  Remission and pancreas isograft survival in recent onset diabetic NOD mice after treatment with low-dose anti-CD3 monoclonal antibodies. , 2002, Transplant immunology.

[111]  R. Elliott,et al.  Immunohistochemical Analyses of Pancreatic Macrophages and CD4 and CD8 T Cell Subsets Prior to and Following Diabetes in the NOD Mouse , 1995, Pancreas.

[112]  R. Elliott,et al.  Distribution of Pancreatic Macrophages Preceding and During Early Insulitis in Young NOD Mice , 1993, Pancreas.

[113]  T. Hanafusa,et al.  Predominance of T lymphocytes in pancreatic islets and spleen of pre-diabetic non-obese diabetic (NOD) mice: a longitudinal study. , 1985, Clinical and experimental immunology.

[114]  R. Darnell,et al.  Dendritic cell maturation is required for the cross-tolerization of CD8+ T cells , 2001, Nature Immunology.

[115]  F. Zhang,et al.  Either IL-2 or IL-12 Is Sufficient to Direct Th1 Differentiation by Nonobese Diabetic T Cells1 , 2003, The Journal of Immunology.

[116]  D. Pardoll,et al.  Cd40-Independent Pathways of T Cell Help for Priming of Cd8+ Cytotoxic T Lymphocytes , 2000, The Journal of experimental medicine.

[117]  F. Pociot,et al.  Onset of type 1 diabetes: a dynamical instability. , 1999, Diabetes.

[118]  P. Kalinski,et al.  Development of Th1-Inducing Capacity in Myeloid Dendritic Cells Requires Environmental Instruction1 , 2000, The Journal of Immunology.

[119]  H. Hall,et al.  CD4+CD25+ regulatory T cells down‐regulate co‐stimulatory molecules on antigen‐presenting cells , 2000, European journal of immunology.

[120]  A. Shapiro,et al.  Combination therapy with sirolimus and interleukin-2 prevents spontaneous and recurrent autoimmune diabetes in NOD mice. , 2002, Diabetes.

[121]  L. Chatenoud,et al.  CD3 antibody-induced dominant self tolerance in overtly diabetic NOD mice. , 1997, Journal of immunology.

[122]  F. Relimpio,et al.  HbA1c levels are better predicted by prebreakfast than postbreakfast blood glucose self-analyses in type 2 diabetes. Influence of duration of diabetes and mode of treatment , 2007, Acta Diabetologica.

[123]  D. Loh,et al.  Antigen-driven differentiation of naive Ig-transgenic B cells in vitro. , 1995, Journal of immunology.

[124]  R. Miura,et al.  A Model of β -Cell Mass, Insulin, and Glucose Kinetics: Pathways to Diabetes , 2000 .

[125]  I. Weissman,et al.  Thymus cell migration: Quantitative aspects of cellular traffic from the thymus to the periphery in mice , 1980, European journal of immunology.

[126]  R. Miura,et al.  A model of beta-cell mass, insulin, and glucose kinetics: pathways to diabetes. , 2000, Journal of theoretical biology.

[127]  Dolca Thomas,et al.  Systemic Transforming Growth Factor-&bgr;1 Gene Therapy Induces Foxp3+ Regulatory Cells, Restores Self-Tolerance, and Facilitates Regeneration Of Beta Cell Function in Overtly Diabetic Nonobese Diabetic Mice , 2005, Transplantation.

[128]  J. Corbett,et al.  Interleukin-1 plus gamma-interferon-induced pancreatic beta-cell dysfunction is mediated by beta-cell nitric oxide production. , 2002, Diabetes.

[129]  A. Lehuen,et al.  NK T Cell-Induced Protection Against Diabetes in Vα14-Jα281 Transgenic Nonobese Diabetic Mice Is Associated with a Th2 Shift Circumscribed Regionally to the Islets and Functionally to Islet Autoantigen1 , 2001, The Journal of Immunology.

[130]  L. Moretta,et al.  NK-dependent DC maturation is mediated by TNFα and IFNγ released upon engagement of the NKp30 triggering receptor , 2005 .

[131]  M. Myers,et al.  Transplacental exposure to bafilomycin disrupts pancreatic islet organogenesis and accelerates diabetes onset in NOD mice. , 2004, Journal of autoimmunity.

[132]  T. Utsugi,et al.  Major Histocompatibility Complex Class I–Restricted Infiltration and Destruction of Pancreatic Islets by NOD Mouse-Derived β-Cell Cytotoxic CD8+ T-Cell Clones In Vivo , 1996, Diabetes.

[133]  D. Wagner,et al.  Peripheral CD 4 loCD 40 + auto-aggressive T cell expansion during insulin-dependent diabetes mellitus , 2004 .

[134]  P. Bedossa,et al.  Role of CD4+CD45RA+ T cells in the development of autoimmune diabetes in the non-obese diabetic (NOD) mouse. , 1993, International immunology.