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Human immune system development and rejection of human islet allografts in spontaneously diabetic NOD-Rag1 IL2rgamma Ins2Akita mice.

Brehm MA, Bortell R, Diiorio P, Leif J, Laning J, Cuthbert A, Yang C, Herlihy M, Burzenski L, Gott B, Foreman O, Powers AC, Greiner DL, Shultz LD - Diabetes (2010)

Bottom Line: To create an immunodeficient mouse model that spontaneously develops hyperglycemia to serve as a diabetic host for human islets and stem cell-derived beta-cells in the absence or presence of a functional human immune system.Engraftment of human hematopoietic stem cells in newborn NRG-Akita and NRG mice resulted in equivalent human immune system development in a normoglycemic or chronically hyperglycemic environment, with >50% of engrafted NRG-Akita mice capable of rejecting human islet allografts.NRG-Akita mice provide a model system for validation of the function of human islets and human adult stem cell, embryonic stem cell, or induced pluripotent stem cell-derived beta-cells in the absence or presence of an alloreactive human immune system.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, USA. michael.brehm@umassmed.edu

ABSTRACT

Objective: To create an immunodeficient mouse model that spontaneously develops hyperglycemia to serve as a diabetic host for human islets and stem cell-derived beta-cells in the absence or presence of a functional human immune system.

Research design and methods: We backcrossed the Ins2(Akita) mutation onto the NOD-Rag1() IL2rgamma() strain and determined 1) the spontaneous development of hyperglycemia, 2) the ability of human islets, mouse islets, and dissociated mouse islet cells to restore euglycemia, 3) the generation of a human immune system following engraftment of human hematopoietic stem cells, and 4) the ability of the humanized mice to reject human islet allografts.

Results: We confirmed the defects in innate and adaptive immunity and the spontaneous development of hyperglycemia conferred by the IL2rgamma(), Rag1(), and Ins2(Akita) genes in NOD-Rag1() IL2rgamma() Ins2(Akita) (NRG-Akita) mice. Mouse and human islets restored NRG-Akita mice to normoglycemia. Insulin-positive cells in dissociated mouse islets, required to restore euglycemia in chemically diabetic NOD-scid IL2rgamma() and spontaneously diabetic NRG-Akita mice, were quantified following transplantation via the intrapancreatic and subrenal routes. Engraftment of human hematopoietic stem cells in newborn NRG-Akita and NRG mice resulted in equivalent human immune system development in a normoglycemic or chronically hyperglycemic environment, with >50% of engrafted NRG-Akita mice capable of rejecting human islet allografts.

Conclusions: NRG-Akita mice provide a model system for validation of the function of human islets and human adult stem cell, embryonic stem cell, or induced pluripotent stem cell-derived beta-cells in the absence or presence of an alloreactive human immune system.

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Transplantation of mouse and human islets into diabetic NRG-Akita mice. Diabetic NRG-Akita mice were transplanted in the renal subcapsular space with 4,000 IEQ human islets or with 20 islets/g body wt mouse islets as described in research design and methods. Blood glucose levels were determined, and the kidney bearing the islet transplant and the host pancreas were recovered at the end of the experiment for histological and immunohistological analyses. A: H-E, insulin, and glucagon staining of transplanted mouse (left panel) and human (right panel) islets and host pancreas of the NRG-Akita transplant recipient at the times indicated after islet transplantation. In the renal subcapsular space, there was robust engraftment of mouse and human islets. Magnification ×200. B: Frequency of diabetes in mouse or human islet recipients. No significant differences were observed between recipients of mouse or human islets. Small vertical bars indicate censored data, i.e., mice that were found dead or were removed from the study for other analyses. (A high-quality digital representation of this figure is available in the online issue.)
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Figure 2: Transplantation of mouse and human islets into diabetic NRG-Akita mice. Diabetic NRG-Akita mice were transplanted in the renal subcapsular space with 4,000 IEQ human islets or with 20 islets/g body wt mouse islets as described in research design and methods. Blood glucose levels were determined, and the kidney bearing the islet transplant and the host pancreas were recovered at the end of the experiment for histological and immunohistological analyses. A: H-E, insulin, and glucagon staining of transplanted mouse (left panel) and human (right panel) islets and host pancreas of the NRG-Akita transplant recipient at the times indicated after islet transplantation. In the renal subcapsular space, there was robust engraftment of mouse and human islets. Magnification ×200. B: Frequency of diabetes in mouse or human islet recipients. No significant differences were observed between recipients of mouse or human islets. Small vertical bars indicate censored data, i.e., mice that were found dead or were removed from the study for other analyses. (A high-quality digital representation of this figure is available in the online issue.)

Mentions: Subrenal transplantation of 20 mouse islets/g body wt into hyperglycemic NRG-Akita mice restored normoglycemia (Fig. 2). Islet graft recipients remained normoglycemic to the end of observation periods or, in selected cases, reverted to hyperglycemia following removal of the graft-bearing kidney (data not shown).


Human immune system development and rejection of human islet allografts in spontaneously diabetic NOD-Rag1 IL2rgamma Ins2Akita mice.

Brehm MA, Bortell R, Diiorio P, Leif J, Laning J, Cuthbert A, Yang C, Herlihy M, Burzenski L, Gott B, Foreman O, Powers AC, Greiner DL, Shultz LD - Diabetes (2010)

Transplantation of mouse and human islets into diabetic NRG-Akita mice. Diabetic NRG-Akita mice were transplanted in the renal subcapsular space with 4,000 IEQ human islets or with 20 islets/g body wt mouse islets as described in research design and methods. Blood glucose levels were determined, and the kidney bearing the islet transplant and the host pancreas were recovered at the end of the experiment for histological and immunohistological analyses. A: H-E, insulin, and glucagon staining of transplanted mouse (left panel) and human (right panel) islets and host pancreas of the NRG-Akita transplant recipient at the times indicated after islet transplantation. In the renal subcapsular space, there was robust engraftment of mouse and human islets. Magnification ×200. B: Frequency of diabetes in mouse or human islet recipients. No significant differences were observed between recipients of mouse or human islets. Small vertical bars indicate censored data, i.e., mice that were found dead or were removed from the study for other analyses. (A high-quality digital representation of this figure is available in the online issue.)
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2927949&req=5

Figure 2: Transplantation of mouse and human islets into diabetic NRG-Akita mice. Diabetic NRG-Akita mice were transplanted in the renal subcapsular space with 4,000 IEQ human islets or with 20 islets/g body wt mouse islets as described in research design and methods. Blood glucose levels were determined, and the kidney bearing the islet transplant and the host pancreas were recovered at the end of the experiment for histological and immunohistological analyses. A: H-E, insulin, and glucagon staining of transplanted mouse (left panel) and human (right panel) islets and host pancreas of the NRG-Akita transplant recipient at the times indicated after islet transplantation. In the renal subcapsular space, there was robust engraftment of mouse and human islets. Magnification ×200. B: Frequency of diabetes in mouse or human islet recipients. No significant differences were observed between recipients of mouse or human islets. Small vertical bars indicate censored data, i.e., mice that were found dead or were removed from the study for other analyses. (A high-quality digital representation of this figure is available in the online issue.)
Mentions: Subrenal transplantation of 20 mouse islets/g body wt into hyperglycemic NRG-Akita mice restored normoglycemia (Fig. 2). Islet graft recipients remained normoglycemic to the end of observation periods or, in selected cases, reverted to hyperglycemia following removal of the graft-bearing kidney (data not shown).

Bottom Line: To create an immunodeficient mouse model that spontaneously develops hyperglycemia to serve as a diabetic host for human islets and stem cell-derived beta-cells in the absence or presence of a functional human immune system.Engraftment of human hematopoietic stem cells in newborn NRG-Akita and NRG mice resulted in equivalent human immune system development in a normoglycemic or chronically hyperglycemic environment, with >50% of engrafted NRG-Akita mice capable of rejecting human islet allografts.NRG-Akita mice provide a model system for validation of the function of human islets and human adult stem cell, embryonic stem cell, or induced pluripotent stem cell-derived beta-cells in the absence or presence of an alloreactive human immune system.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, USA. michael.brehm@umassmed.edu

ABSTRACT

Objective: To create an immunodeficient mouse model that spontaneously develops hyperglycemia to serve as a diabetic host for human islets and stem cell-derived beta-cells in the absence or presence of a functional human immune system.

Research design and methods: We backcrossed the Ins2(Akita) mutation onto the NOD-Rag1() IL2rgamma() strain and determined 1) the spontaneous development of hyperglycemia, 2) the ability of human islets, mouse islets, and dissociated mouse islet cells to restore euglycemia, 3) the generation of a human immune system following engraftment of human hematopoietic stem cells, and 4) the ability of the humanized mice to reject human islet allografts.

Results: We confirmed the defects in innate and adaptive immunity and the spontaneous development of hyperglycemia conferred by the IL2rgamma(), Rag1(), and Ins2(Akita) genes in NOD-Rag1() IL2rgamma() Ins2(Akita) (NRG-Akita) mice. Mouse and human islets restored NRG-Akita mice to normoglycemia. Insulin-positive cells in dissociated mouse islets, required to restore euglycemia in chemically diabetic NOD-scid IL2rgamma() and spontaneously diabetic NRG-Akita mice, were quantified following transplantation via the intrapancreatic and subrenal routes. Engraftment of human hematopoietic stem cells in newborn NRG-Akita and NRG mice resulted in equivalent human immune system development in a normoglycemic or chronically hyperglycemic environment, with >50% of engrafted NRG-Akita mice capable of rejecting human islet allografts.

Conclusions: NRG-Akita mice provide a model system for validation of the function of human islets and human adult stem cell, embryonic stem cell, or induced pluripotent stem cell-derived beta-cells in the absence or presence of an alloreactive human immune system.

Show MeSH
Related in: MedlinePlus