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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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Phenotype and functional activity of CD8+ T cells from RAG-2−/− versus RAG-2+ 8.3-NOD mice. (A) Flow cytometry profiles of  CD4+CD8+ (left) and CD4−CD8+ thymocytes (right) from 8.3-NOD mice (dotted line) and RAG-2−/− 8.3-NOD mice (solid line) for maturation markers. Thymocytes were analyzed by three-color cytofluorometry as in Fig. 1. Panels show the fluorescence histograms of each marker on gated  CD4+CD8+ and CD4−CD8+ thymocytes. (B) Flow cytometry profiles of splenic CD8+ T cells from 8.3-NOD mice (dotted line) and RAG-2−/− 8.3-NOD mice (solid line) for activation and memory markers. (C) Proliferative activity of splenic CD8+ T cells from 8.3-NOD mice and RAG-2−/− 8.3-NOD mice in response to islet cells. (D) Reverse transcription-PCR analysis for cytokine gene expression of islet-derived CD8+ T cells from 8.3-NOD  mice and RAG-2−/− 8.3-NOD mice. M, 1Kb ladder. (E) Kinetics of insulitis in RAG-2−/− 8.3-NOD mice.
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Figure 5: Phenotype and functional activity of CD8+ T cells from RAG-2−/− versus RAG-2+ 8.3-NOD mice. (A) Flow cytometry profiles of CD4+CD8+ (left) and CD4−CD8+ thymocytes (right) from 8.3-NOD mice (dotted line) and RAG-2−/− 8.3-NOD mice (solid line) for maturation markers. Thymocytes were analyzed by three-color cytofluorometry as in Fig. 1. Panels show the fluorescence histograms of each marker on gated CD4+CD8+ and CD4−CD8+ thymocytes. (B) Flow cytometry profiles of splenic CD8+ T cells from 8.3-NOD mice (dotted line) and RAG-2−/− 8.3-NOD mice (solid line) for activation and memory markers. (C) Proliferative activity of splenic CD8+ T cells from 8.3-NOD mice and RAG-2−/− 8.3-NOD mice in response to islet cells. (D) Reverse transcription-PCR analysis for cytokine gene expression of islet-derived CD8+ T cells from 8.3-NOD mice and RAG-2−/− 8.3-NOD mice. M, 1Kb ladder. (E) Kinetics of insulitis in RAG-2−/− 8.3-NOD mice.

Mentions: Experiments were next performed to elucidate the mechanisms underlying disease deceleration in RAG-2−/− 8.3-NOD versus RAG-2+ 8.3-NOD mice. Three-color cytofluorometric studies using mAbs against several differentiation markers, including the transgenic TCR, MHC class I (Kd), CD5, CD24, CD44, and CD69, revealed that the CD4+CD8+ and CD4−CD8+ thymocytes of RAG-2−/− 8.3-NOD mice and RAG-2+ 8.3-NOD mice were phenotypically similar (Fig. 5 A). Likewise, no phenotypic differences were noted between the peripheral CD8+ T cells of RAG-2−/− 8.3-NOD mice and those of RAG-2+ 8.3-NOD mice with respect to several markers, including Vβ8.1/8.2, CD2, CD11a, CD28, CD44, CD45RB, CD62L and CD69 (Fig. 5 B). Furthermore, splenic CD8+ T cells from both types of mice proliferated equally well in response to islet cells in vitro (Fig. 5 C). It thus appears that the 8.3-CD8+ T cells of RAG-2−/− 8.3-NOD mice are phenotypically and functionally similar to those of RAG-2+ 8.3-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)

Phenotype and functional activity of CD8+ T cells from RAG-2−/− versus RAG-2+ 8.3-NOD mice. (A) Flow cytometry profiles of  CD4+CD8+ (left) and CD4−CD8+ thymocytes (right) from 8.3-NOD mice (dotted line) and RAG-2−/− 8.3-NOD mice (solid line) for maturation markers. Thymocytes were analyzed by three-color cytofluorometry as in Fig. 1. Panels show the fluorescence histograms of each marker on gated  CD4+CD8+ and CD4−CD8+ thymocytes. (B) Flow cytometry profiles of splenic CD8+ T cells from 8.3-NOD mice (dotted line) and RAG-2−/− 8.3-NOD mice (solid line) for activation and memory markers. (C) Proliferative activity of splenic CD8+ T cells from 8.3-NOD mice and RAG-2−/− 8.3-NOD mice in response to islet cells. (D) Reverse transcription-PCR analysis for cytokine gene expression of islet-derived CD8+ T cells from 8.3-NOD  mice and RAG-2−/− 8.3-NOD mice. M, 1Kb ladder. (E) Kinetics of insulitis in RAG-2−/− 8.3-NOD mice.
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Figure 5: Phenotype and functional activity of CD8+ T cells from RAG-2−/− versus RAG-2+ 8.3-NOD mice. (A) Flow cytometry profiles of CD4+CD8+ (left) and CD4−CD8+ thymocytes (right) from 8.3-NOD mice (dotted line) and RAG-2−/− 8.3-NOD mice (solid line) for maturation markers. Thymocytes were analyzed by three-color cytofluorometry as in Fig. 1. Panels show the fluorescence histograms of each marker on gated CD4+CD8+ and CD4−CD8+ thymocytes. (B) Flow cytometry profiles of splenic CD8+ T cells from 8.3-NOD mice (dotted line) and RAG-2−/− 8.3-NOD mice (solid line) for activation and memory markers. (C) Proliferative activity of splenic CD8+ T cells from 8.3-NOD mice and RAG-2−/− 8.3-NOD mice in response to islet cells. (D) Reverse transcription-PCR analysis for cytokine gene expression of islet-derived CD8+ T cells from 8.3-NOD mice and RAG-2−/− 8.3-NOD mice. M, 1Kb ladder. (E) Kinetics of insulitis in RAG-2−/− 8.3-NOD mice.
Mentions: Experiments were next performed to elucidate the mechanisms underlying disease deceleration in RAG-2−/− 8.3-NOD versus RAG-2+ 8.3-NOD mice. Three-color cytofluorometric studies using mAbs against several differentiation markers, including the transgenic TCR, MHC class I (Kd), CD5, CD24, CD44, and CD69, revealed that the CD4+CD8+ and CD4−CD8+ thymocytes of RAG-2−/− 8.3-NOD mice and RAG-2+ 8.3-NOD mice were phenotypically similar (Fig. 5 A). Likewise, no phenotypic differences were noted between the peripheral CD8+ T cells of RAG-2−/− 8.3-NOD mice and those of RAG-2+ 8.3-NOD mice with respect to several markers, including Vβ8.1/8.2, CD2, CD11a, CD28, CD44, CD45RB, CD62L and CD69 (Fig. 5 B). Furthermore, splenic CD8+ T cells from both types of mice proliferated equally well in response to islet cells in vitro (Fig. 5 C). It thus appears that the 8.3-CD8+ T cells of RAG-2−/− 8.3-NOD mice are phenotypically and functionally similar to those of RAG-2+ 8.3-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