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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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Expression of the TCR-α/β transgenes in 8.3-NOD mice. CD4, CD8, and Vβ8.1/8.2 profiles of thymocytes (A) and lymph node cells (B)  from nontransgenic NOD, 8.3–TCR-β–transgenic NOD, and 8.3-NOD mice. (Top) CD4 versus CD8 contour plots of cell suspensions stained with  anti-CD8-PE, anti-V β8.1/8.2-FITC, and anti-CD4-biotin plus Streptavidin-PerCP. The lower panels show the Vβ8.1/8.2 fluorescence histograms of  each T cell subset after electronic gating. Numbers indicate the average percentage of cells (top) or the average number of Vβ8.1/8.2+ cells (bottom) in  each subset. Data correspond to 3–9 mice/group. DP, double-positive cells; DN, double-negative cells.
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Figure 1: Expression of the TCR-α/β transgenes in 8.3-NOD mice. CD4, CD8, and Vβ8.1/8.2 profiles of thymocytes (A) and lymph node cells (B) from nontransgenic NOD, 8.3–TCR-β–transgenic NOD, and 8.3-NOD mice. (Top) CD4 versus CD8 contour plots of cell suspensions stained with anti-CD8-PE, anti-V β8.1/8.2-FITC, and anti-CD4-biotin plus Streptavidin-PerCP. The lower panels show the Vβ8.1/8.2 fluorescence histograms of each T cell subset after electronic gating. Numbers indicate the average percentage of cells (top) or the average number of Vβ8.1/8.2+ cells (bottom) in each subset. Data correspond to 3–9 mice/group. DP, double-positive cells; DN, double-negative cells.

Mentions: Three-color cytofluorometric studies showed that >97% CD4−CD8+ thymocytes (Fig. 1 A) and lymph node CD8+ T cells (Fig. 1 B) from 8.3-NOD mice expressed Vβ8.1/8.2+ TCRs, as compared with 18% of the CD4−CD8+ thymocytes and lymph node CD8+ T cells from nontransgenic NOD mice, indicating TCR-β transgene expression. Although we do not have a TCR-α–transgenic chain–specific mAb, and thus cannot directly quantitate its expression, three different lines of evidence indicate that the transgenic TCR-α chain is expressed on a significant fraction of thymocytes and peripheral T cells from 8.3-NOD mice. First, ∼67% of CD4+CD8+ thymocytes from 8.3-NOD mice expressed high levels of the transgenic TCR-β chain, as compared to only 18% of CD4+CD8+ thymocytes from TCR-β–transgenic NOD mice, suggesting early TCR-α chain expression and early upregulation of TCR-α/β heterodimers on immature thymocytes (38–42). Second, thymocyte development in 8.3-NOD mice, but not 8.3–TCR-β–transgenic NOD mice, was skewed towards the CD4−CD8+ subset (Fig. 1 A), suggesting TCR-α–transgene–dependent positive selection of 8.3-CD8+ thymocytes as in other MHC class I–restricted TCR-α/β–transgenic models (38, 41, 43). As a result, the lymph nodes of 8.3-NOD mice consistently had many more CD8+ T cells (>65%) and fewer CD4+ T cells (<15%) than the lymph nodes of 8.3– TCR-β–transgenic and nontransgenic NOD mice (with <25% CD8+ T cells and >40% CD4+ T cells) (Fig. 1 B). Finally, introduction of the 8.3–TCR-α and –TCR-β transgenes into RAG-2−/− NOD mice, which cannot rearrange endogenous TCR genes, resulted in the positive selection of CD8+, but not CD4+, Vβ8.1/8.2+ T cells (see below). Taken together, these data indicate that the 8.3-TCR is expressed on a significant fraction of thymocytes and peripheral T cells from 8.3-NOD mice, and that 8.3– TCR-α/β–transgene expression fosters the positive selection of CD8+ T cells.


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)

Expression of the TCR-α/β transgenes in 8.3-NOD mice. CD4, CD8, and Vβ8.1/8.2 profiles of thymocytes (A) and lymph node cells (B)  from nontransgenic NOD, 8.3–TCR-β–transgenic NOD, and 8.3-NOD mice. (Top) CD4 versus CD8 contour plots of cell suspensions stained with  anti-CD8-PE, anti-V β8.1/8.2-FITC, and anti-CD4-biotin plus Streptavidin-PerCP. The lower panels show the Vβ8.1/8.2 fluorescence histograms of  each T cell subset after electronic gating. Numbers indicate the average percentage of cells (top) or the average number of Vβ8.1/8.2+ cells (bottom) in  each subset. Data correspond to 3–9 mice/group. DP, double-positive cells; DN, double-negative cells.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2199139&req=5

Figure 1: Expression of the TCR-α/β transgenes in 8.3-NOD mice. CD4, CD8, and Vβ8.1/8.2 profiles of thymocytes (A) and lymph node cells (B) from nontransgenic NOD, 8.3–TCR-β–transgenic NOD, and 8.3-NOD mice. (Top) CD4 versus CD8 contour plots of cell suspensions stained with anti-CD8-PE, anti-V β8.1/8.2-FITC, and anti-CD4-biotin plus Streptavidin-PerCP. The lower panels show the Vβ8.1/8.2 fluorescence histograms of each T cell subset after electronic gating. Numbers indicate the average percentage of cells (top) or the average number of Vβ8.1/8.2+ cells (bottom) in each subset. Data correspond to 3–9 mice/group. DP, double-positive cells; DN, double-negative cells.
Mentions: Three-color cytofluorometric studies showed that >97% CD4−CD8+ thymocytes (Fig. 1 A) and lymph node CD8+ T cells (Fig. 1 B) from 8.3-NOD mice expressed Vβ8.1/8.2+ TCRs, as compared with 18% of the CD4−CD8+ thymocytes and lymph node CD8+ T cells from nontransgenic NOD mice, indicating TCR-β transgene expression. Although we do not have a TCR-α–transgenic chain–specific mAb, and thus cannot directly quantitate its expression, three different lines of evidence indicate that the transgenic TCR-α chain is expressed on a significant fraction of thymocytes and peripheral T cells from 8.3-NOD mice. First, ∼67% of CD4+CD8+ thymocytes from 8.3-NOD mice expressed high levels of the transgenic TCR-β chain, as compared to only 18% of CD4+CD8+ thymocytes from TCR-β–transgenic NOD mice, suggesting early TCR-α chain expression and early upregulation of TCR-α/β heterodimers on immature thymocytes (38–42). Second, thymocyte development in 8.3-NOD mice, but not 8.3–TCR-β–transgenic NOD mice, was skewed towards the CD4−CD8+ subset (Fig. 1 A), suggesting TCR-α–transgene–dependent positive selection of 8.3-CD8+ thymocytes as in other MHC class I–restricted TCR-α/β–transgenic models (38, 41, 43). As a result, the lymph nodes of 8.3-NOD mice consistently had many more CD8+ T cells (>65%) and fewer CD4+ T cells (<15%) than the lymph nodes of 8.3– TCR-β–transgenic and nontransgenic NOD mice (with <25% CD8+ T cells and >40% CD4+ T cells) (Fig. 1 B). Finally, introduction of the 8.3–TCR-α and –TCR-β transgenes into RAG-2−/− NOD mice, which cannot rearrange endogenous TCR genes, resulted in the positive selection of CD8+, but not CD4+, Vβ8.1/8.2+ T cells (see below). Taken together, these data indicate that the 8.3-TCR is expressed on a significant fraction of thymocytes and peripheral T cells from 8.3-NOD mice, and that 8.3– TCR-α/β–transgene expression fosters the positive selection of CD8+ T cells.

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