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Spontaneous autoimmune diabetes in monoclonal T cell nonobese diabetic mice.

Verdaguer J, Schmidt D, Amrani A, Anderson B, Averill N, Santamaria P - J. Exp. Med. (1997)

Bottom Line: The precise roles that beta cell-reactive CD8+ and CD4+ T cells play in the disease process, however, remain ill defined.Here we have investigated whether naive beta cell-specific CD8+ and CD4+ T cells can spontaneously accumulate in pancreatic islets, differentiate into effector cells, and destroy beta cells in the absence of other T cell specificities.These results demonstrate that naive beta cell- specific CD8+ and CD4+ T cells can trigger diabetes in the absence of other T or B cell specificities, but suggest that efficient recruitment of naive diabetogenic beta cell-reactive CD8+ T cells to islets requires the assistance of beta cell-reactive CD4+ T cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Infectious Diseases, The University of Calgary, Faculty of Medicine, Alberta, Canada.

ABSTRACT
It has been established that insulin-dependent diabetes mellitus (IDDM) in nonobese diabetic (NOD) mice results from a CD4+ and CD8+ T cell-dependent autoimmune process directed against the pancreatic beta cells. The precise roles that beta cell-reactive CD8+ and CD4+ T cells play in the disease process, however, remain ill defined. Here we have investigated whether naive beta cell-specific CD8+ and CD4+ T cells can spontaneously accumulate in pancreatic islets, differentiate into effector cells, and destroy beta cells in the absence of other T cell specificities. This was done by introducing Kd- or I-Ag7-restricted beta cell-specific T cell receptor (TCR) transgenes that are highly diabetogenic in NOD mice (8.3- and 4.1-TCR, respectively), into recombination-activating gene (RAG)-2-deficient NOD mice, which cannot rearrange endogenous TCR genes and thus bear monoclonal TCR repertoires. We show that while RAG-2(-/-) 4.1-NOD mice, which only bear beta cell-specific CD4+ T cells, develop diabetes as early and as frequently as RAG-2+ 4.1-NOD mice, RAG-2(-/-) 8.3-NOD mice, which only bear beta cell-specific CD8+ T cells, develop diabetes less frequently and significantly later than RAG-2(+) 8.3-NOD mice. The monoclonal CD8+ T cells of RAG-2(-/-) 8.3-NOD mice mature properly, proliferate vigorously in response to antigenic stimulation in vitro, and can differentiate into beta cell-cytotoxic T cells in vivo, but do not efficiently accumulate in islets in the absence of a CD4+ T cell-derived signal, which can be provided by splenic CD4+ T cells from nontransgenic NOD mice. These results demonstrate that naive beta cell- specific CD8+ and CD4+ T cells can trigger diabetes in the absence of other T or B cell specificities, but suggest that efficient recruitment of naive diabetogenic beta cell-reactive CD8+ T cells to islets requires the assistance of beta cell-reactive CD4+ T cells.

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8.3–TCR-α/β– transgene expression and diabetogenesis. (A) Incidence of  IDDM in female (26 8.3–, 21  8.3–TCR-β–transgenic, and 114  nontransgenic) and male (24  8.3–, 30 8.3–TCR-β–transgenic, and 59 nontransgenic)  NOD mice. (B) Progression of  insulitis in nontransgenic, 8.3– TCR-β–transgenic, and 8.3-NOD mice (4–7 mice/age group;  15–30 islets/mouse). Bars show  the standard deviation of the  means. *, P <0.0001 (Mann-Whitney U test). (C) Flow cytometry profile of islet-derived T  cells from diabetic 8.3-NOD  mice. Islets from acutely diabetic  8.3-NOD mice were cultured in  rIL-2–containing CM for 3–5 d  and the resulting cells studied by  flow cytometry as in Fig. 1. (D)  Phenotype of islet-infiltrating T  cells in 8.3-NOD versus nontransgenic NOD mice. Pancreas  sections were stained with anti-CD8 (53.6-7) or anti-CD4  (GK1.5) mAbs and FITC-labeled  anti–rat IgG, and observed under  a fluorescence microscope. Original magnification: 200.
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Figure 3: 8.3–TCR-α/β– transgene expression and diabetogenesis. (A) Incidence of IDDM in female (26 8.3–, 21 8.3–TCR-β–transgenic, and 114 nontransgenic) and male (24 8.3–, 30 8.3–TCR-β–transgenic, and 59 nontransgenic) NOD mice. (B) Progression of insulitis in nontransgenic, 8.3– TCR-β–transgenic, and 8.3-NOD mice (4–7 mice/age group; 15–30 islets/mouse). Bars show the standard deviation of the means. *, P <0.0001 (Mann-Whitney U test). (C) Flow cytometry profile of islet-derived T cells from diabetic 8.3-NOD mice. Islets from acutely diabetic 8.3-NOD mice were cultured in rIL-2–containing CM for 3–5 d and the resulting cells studied by flow cytometry as in Fig. 1. (D) Phenotype of islet-infiltrating T cells in 8.3-NOD versus nontransgenic NOD mice. Pancreas sections were stained with anti-CD8 (53.6-7) or anti-CD4 (GK1.5) mAbs and FITC-labeled anti–rat IgG, and observed under a fluorescence microscope. Original magnification: 200.

Mentions: We next investigated whether positive selection of the beta cell–specific 8.3-TCR in 8.3-NOD mice had any pathogenic significance. We therefore followed 8.3-NOD mice of the N3-N6 backcrosses of the founder mice onto the NOD background for development of diabetes, and compared the cumulative incidence curves of these mice with those of nontransgenic NOD mice and 8.3–TCR-β–transgenic NOD mice (26). As shown in Fig. 3 A, female and male 8.3-NOD mice developed diabetes much earlier than female and male 8.3–TCR-β–transgenic (N4-N7), nontransgenic NOD littermates (N3-N5) or NOD/Lt mice (ages at onset: 43 ± 10 versus 88 ± 12, 120 ± 19 and 119 ± 26 d, respectively, for females, P <0.0001; and 66 ± 15 versus 109 ± 33, 173 ± 35, and 157 ± 28 d, respectively, for males, P <0.0001). Despite the dramatic acceleration of diabetes onset in 8.3-NOD mice, the cumulative incidence of diabetes and the kinetics of disease penetrance in the 8.3-NOD, 8.3–TCR-β–transgenic, nontransgenic NOD littermates, and NOD/ Lt mouse populations were remarkably similar (females: 73.1% versus 86, 78, and 84%, respectively; males: 50% versus 50, 31, and 46%, respectively; Fig. 3 A). No major differences in disease penetrance were noted among 8.3-NOD mice of the N3-N6 generations. In females, for example, the incidence and age at onset of diabetes for each generation were: N3, 6/8 mice at 40 ± 4 d; N4, 5/6 mice at 45 ± 6 d; N5, 2/3 mice at 37 ± 3 d; and N6, 6/9 mice at 51 ± 13 d. The same was true for 8.3–TCR-β–transgenic NOD mice (26), and for nontransgenic NOD littermates (N3 females: 3/3 mice at 140 ± 5 d; N4 females: 6/ 8 mice at 117 ± 46 d; and N5 females: 5/7 mice at 117 ± 14 d). Taken together, these data indicate that the 8.3-TCR is highly diabetogenic in the NOD background, and suggest that the events that trigger diabetes in 8.3-NOD mice are similar to those that trigger diabetes in nontransgenic NOD mice.


Spontaneous autoimmune diabetes in monoclonal T cell nonobese diabetic mice.

Verdaguer J, Schmidt D, Amrani A, Anderson B, Averill N, Santamaria P - J. Exp. Med. (1997)

8.3–TCR-α/β– transgene expression and diabetogenesis. (A) Incidence of  IDDM in female (26 8.3–, 21  8.3–TCR-β–transgenic, and 114  nontransgenic) and male (24  8.3–, 30 8.3–TCR-β–transgenic, and 59 nontransgenic)  NOD mice. (B) Progression of  insulitis in nontransgenic, 8.3– TCR-β–transgenic, and 8.3-NOD mice (4–7 mice/age group;  15–30 islets/mouse). Bars show  the standard deviation of the  means. *, P <0.0001 (Mann-Whitney U test). (C) Flow cytometry profile of islet-derived T  cells from diabetic 8.3-NOD  mice. Islets from acutely diabetic  8.3-NOD mice were cultured in  rIL-2–containing CM for 3–5 d  and the resulting cells studied by  flow cytometry as in Fig. 1. (D)  Phenotype of islet-infiltrating T  cells in 8.3-NOD versus nontransgenic NOD mice. Pancreas  sections were stained with anti-CD8 (53.6-7) or anti-CD4  (GK1.5) mAbs and FITC-labeled  anti–rat IgG, and observed under  a fluorescence microscope. Original magnification: 200.
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Related In: Results  -  Collection

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Figure 3: 8.3–TCR-α/β– transgene expression and diabetogenesis. (A) Incidence of IDDM in female (26 8.3–, 21 8.3–TCR-β–transgenic, and 114 nontransgenic) and male (24 8.3–, 30 8.3–TCR-β–transgenic, and 59 nontransgenic) NOD mice. (B) Progression of insulitis in nontransgenic, 8.3– TCR-β–transgenic, and 8.3-NOD mice (4–7 mice/age group; 15–30 islets/mouse). Bars show the standard deviation of the means. *, P <0.0001 (Mann-Whitney U test). (C) Flow cytometry profile of islet-derived T cells from diabetic 8.3-NOD mice. Islets from acutely diabetic 8.3-NOD mice were cultured in rIL-2–containing CM for 3–5 d and the resulting cells studied by flow cytometry as in Fig. 1. (D) Phenotype of islet-infiltrating T cells in 8.3-NOD versus nontransgenic NOD mice. Pancreas sections were stained with anti-CD8 (53.6-7) or anti-CD4 (GK1.5) mAbs and FITC-labeled anti–rat IgG, and observed under a fluorescence microscope. Original magnification: 200.
Mentions: We next investigated whether positive selection of the beta cell–specific 8.3-TCR in 8.3-NOD mice had any pathogenic significance. We therefore followed 8.3-NOD mice of the N3-N6 backcrosses of the founder mice onto the NOD background for development of diabetes, and compared the cumulative incidence curves of these mice with those of nontransgenic NOD mice and 8.3–TCR-β–transgenic NOD mice (26). As shown in Fig. 3 A, female and male 8.3-NOD mice developed diabetes much earlier than female and male 8.3–TCR-β–transgenic (N4-N7), nontransgenic NOD littermates (N3-N5) or NOD/Lt mice (ages at onset: 43 ± 10 versus 88 ± 12, 120 ± 19 and 119 ± 26 d, respectively, for females, P <0.0001; and 66 ± 15 versus 109 ± 33, 173 ± 35, and 157 ± 28 d, respectively, for males, P <0.0001). Despite the dramatic acceleration of diabetes onset in 8.3-NOD mice, the cumulative incidence of diabetes and the kinetics of disease penetrance in the 8.3-NOD, 8.3–TCR-β–transgenic, nontransgenic NOD littermates, and NOD/ Lt mouse populations were remarkably similar (females: 73.1% versus 86, 78, and 84%, respectively; males: 50% versus 50, 31, and 46%, respectively; Fig. 3 A). No major differences in disease penetrance were noted among 8.3-NOD mice of the N3-N6 generations. In females, for example, the incidence and age at onset of diabetes for each generation were: N3, 6/8 mice at 40 ± 4 d; N4, 5/6 mice at 45 ± 6 d; N5, 2/3 mice at 37 ± 3 d; and N6, 6/9 mice at 51 ± 13 d. The same was true for 8.3–TCR-β–transgenic NOD mice (26), and for nontransgenic NOD littermates (N3 females: 3/3 mice at 140 ± 5 d; N4 females: 6/ 8 mice at 117 ± 46 d; and N5 females: 5/7 mice at 117 ± 14 d). Taken together, these data indicate that the 8.3-TCR is highly diabetogenic in the NOD background, and suggest that the events that trigger diabetes in 8.3-NOD mice are similar to those that trigger diabetes in nontransgenic NOD mice.

Bottom Line: The precise roles that beta cell-reactive CD8+ and CD4+ T cells play in the disease process, however, remain ill defined.Here we have investigated whether naive beta cell-specific CD8+ and CD4+ T cells can spontaneously accumulate in pancreatic islets, differentiate into effector cells, and destroy beta cells in the absence of other T cell specificities.These results demonstrate that naive beta cell- specific CD8+ and CD4+ T cells can trigger diabetes in the absence of other T or B cell specificities, but suggest that efficient recruitment of naive diabetogenic beta cell-reactive CD8+ T cells to islets requires the assistance of beta cell-reactive CD4+ T cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Infectious Diseases, The University of Calgary, Faculty of Medicine, Alberta, Canada.

ABSTRACT
It has been established that insulin-dependent diabetes mellitus (IDDM) in nonobese diabetic (NOD) mice results from a CD4+ and CD8+ T cell-dependent autoimmune process directed against the pancreatic beta cells. The precise roles that beta cell-reactive CD8+ and CD4+ T cells play in the disease process, however, remain ill defined. Here we have investigated whether naive beta cell-specific CD8+ and CD4+ T cells can spontaneously accumulate in pancreatic islets, differentiate into effector cells, and destroy beta cells in the absence of other T cell specificities. This was done by introducing Kd- or I-Ag7-restricted beta cell-specific T cell receptor (TCR) transgenes that are highly diabetogenic in NOD mice (8.3- and 4.1-TCR, respectively), into recombination-activating gene (RAG)-2-deficient NOD mice, which cannot rearrange endogenous TCR genes and thus bear monoclonal TCR repertoires. We show that while RAG-2(-/-) 4.1-NOD mice, which only bear beta cell-specific CD4+ T cells, develop diabetes as early and as frequently as RAG-2+ 4.1-NOD mice, RAG-2(-/-) 8.3-NOD mice, which only bear beta cell-specific CD8+ T cells, develop diabetes less frequently and significantly later than RAG-2(+) 8.3-NOD mice. The monoclonal CD8+ T cells of RAG-2(-/-) 8.3-NOD mice mature properly, proliferate vigorously in response to antigenic stimulation in vitro, and can differentiate into beta cell-cytotoxic T cells in vivo, but do not efficiently accumulate in islets in the absence of a CD4+ T cell-derived signal, which can be provided by splenic CD4+ T cells from nontransgenic NOD mice. These results demonstrate that naive beta cell- specific CD8+ and CD4+ T cells can trigger diabetes in the absence of other T or B cell specificities, but suggest that efficient recruitment of naive diabetogenic beta cell-reactive CD8+ T cells to islets requires the assistance of beta cell-reactive CD4+ T cells.

Show MeSH
Related in: MedlinePlus