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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 human islet allografts into diabetic NRG-Akita mice engrafted with human HSC. Diabetic NRG-Akita and HSC-engrafted NRG-Akita mice were transplanted subrenally with 4,000 human IEQ as described in research design and methods. A: Frequency of diabetes in islet allograft recipients. NRG-Akita vs. HSC-engrafted NRG-Akita, P = 0.03. B: Representative histology and immunochemical staining patterns are shown. Note the abundance of insulin-positive cells and the absence of human CD45-positive cells in non–HSC engrafted NRG-Akita mice (left panel). Note the presence of fewer insulin-positive cells and islet graft infiltration by human CD45+ cells in HSC-engrafted NRG-Akita mice that were normoglycemic at the end of the experiment (middle panel). Note the scarcity of insulin-positive cells and moderate numbers of human CD45+ cells in HSC-engrafted NRG Akita mice that were hyperglycemic at the end of the experiment and had rejected their human islet allografts (right panel). Magnification ×200. huCD45, human CD45 staining; Insulin, insulin staining. (A high-quality digital representation of this figure is available in the online issue.)
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Figure 3: Transplantation of human islet allografts into diabetic NRG-Akita mice engrafted with human HSC. Diabetic NRG-Akita and HSC-engrafted NRG-Akita mice were transplanted subrenally with 4,000 human IEQ as described in research design and methods. A: Frequency of diabetes in islet allograft recipients. NRG-Akita vs. HSC-engrafted NRG-Akita, P = 0.03. B: Representative histology and immunochemical staining patterns are shown. Note the abundance of insulin-positive cells and the absence of human CD45-positive cells in non–HSC engrafted NRG-Akita mice (left panel). Note the presence of fewer insulin-positive cells and islet graft infiltration by human CD45+ cells in HSC-engrafted NRG-Akita mice that were normoglycemic at the end of the experiment (middle panel). Note the scarcity of insulin-positive cells and moderate numbers of human CD45+ cells in HSC-engrafted NRG Akita mice that were hyperglycemic at the end of the experiment and had rejected their human islet allografts (right panel). Magnification ×200. huCD45, human CD45 staining; Insulin, insulin staining. (A high-quality digital representation of this figure is available in the online issue.)

Mentions: Eight of nine human islet recipients of 4,000 IEQ restored normoglycemia (<250 mg/dl) (Fig. 3) in non–HSC-engrafted diabetic NRG-Akita mice. In contrast, eight of 13 human islet allografts were rejected in HSC-engrafted NRG-Akita mice (Fig. 3). Insulin staining in the absence of mononuclear cell infiltration was detected in human islet allografts in non–HSC-engrafted NRG-Akita mice (Fig. 3). In the eight HSC-engrafted NRG-Akita human islet recipients that reverted to hyperglycemia, human CD45+ cell infiltration into the graft site and loss of β-cells were observed. In the five HSC-engrafted NRG-Akita mice that did not revert to hyperglycemia, insulin-positive cells and human CD45+ cell islet infiltration were observed (Fig. 3).


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 human islet allografts into diabetic NRG-Akita mice engrafted with human HSC. Diabetic NRG-Akita and HSC-engrafted NRG-Akita mice were transplanted subrenally with 4,000 human IEQ as described in research design and methods. A: Frequency of diabetes in islet allograft recipients. NRG-Akita vs. HSC-engrafted NRG-Akita, P = 0.03. B: Representative histology and immunochemical staining patterns are shown. Note the abundance of insulin-positive cells and the absence of human CD45-positive cells in non–HSC engrafted NRG-Akita mice (left panel). Note the presence of fewer insulin-positive cells and islet graft infiltration by human CD45+ cells in HSC-engrafted NRG-Akita mice that were normoglycemic at the end of the experiment (middle panel). Note the scarcity of insulin-positive cells and moderate numbers of human CD45+ cells in HSC-engrafted NRG Akita mice that were hyperglycemic at the end of the experiment and had rejected their human islet allografts (right panel). Magnification ×200. huCD45, human CD45 staining; Insulin, insulin staining. (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 3: Transplantation of human islet allografts into diabetic NRG-Akita mice engrafted with human HSC. Diabetic NRG-Akita and HSC-engrafted NRG-Akita mice were transplanted subrenally with 4,000 human IEQ as described in research design and methods. A: Frequency of diabetes in islet allograft recipients. NRG-Akita vs. HSC-engrafted NRG-Akita, P = 0.03. B: Representative histology and immunochemical staining patterns are shown. Note the abundance of insulin-positive cells and the absence of human CD45-positive cells in non–HSC engrafted NRG-Akita mice (left panel). Note the presence of fewer insulin-positive cells and islet graft infiltration by human CD45+ cells in HSC-engrafted NRG-Akita mice that were normoglycemic at the end of the experiment (middle panel). Note the scarcity of insulin-positive cells and moderate numbers of human CD45+ cells in HSC-engrafted NRG Akita mice that were hyperglycemic at the end of the experiment and had rejected their human islet allografts (right panel). Magnification ×200. huCD45, human CD45 staining; Insulin, insulin staining. (A high-quality digital representation of this figure is available in the online issue.)
Mentions: Eight of nine human islet recipients of 4,000 IEQ restored normoglycemia (<250 mg/dl) (Fig. 3) in non–HSC-engrafted diabetic NRG-Akita mice. In contrast, eight of 13 human islet allografts were rejected in HSC-engrafted NRG-Akita mice (Fig. 3). Insulin staining in the absence of mononuclear cell infiltration was detected in human islet allografts in non–HSC-engrafted NRG-Akita mice (Fig. 3). In the eight HSC-engrafted NRG-Akita human islet recipients that reverted to hyperglycemia, human CD45+ cell infiltration into the graft site and loss of β-cells were observed. In the five HSC-engrafted NRG-Akita mice that did not revert to hyperglycemia, insulin-positive cells and human CD45+ cell islet infiltration were observed (Fig. 3).

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