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CD4(+) T cells from glutamic acid decarboxylase (GAD)65-specific T cell receptor transgenic mice are not diabetogenic and can delay diabetes transfer.

Tarbell KV, Lee M, Ranheim E, Chao CC, Sanna M, Kim SK, Dickie P, Teyton L, Davis M, McDevitt H - J. Exp. Med. (2002)

Bottom Line: However, the exact role of GAD65-specific T cells in diabetes pathogenesis is unclear.Lymphocytes from these TCR transgenic mice proliferate and make interferon gamma, interleukin (IL)-2, tumor necrosis factor (TNF)-alpha, and IL-10 when stimulated in vitro with GAD65 peptide 286-300, yet these TCR transgenic animals do not spontaneously develop diabetes, and insulitis is virtually undetectable.This data suggests that GAD65 peptide 286-300-specific T cells have disease protective capacity and are not pathogenic.

View Article: PubMed Central - PubMed

Affiliation: Program in Immunology, Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA.

ABSTRACT
Glutamic acid decarboxylase (GAD)65 is an early and important antigen in both human diabetes mellitus and the nonobese diabetic (NOD) mouse. However, the exact role of GAD65-specific T cells in diabetes pathogenesis is unclear. T cell responses to GAD65 occur early in diabetes pathogenesis, yet only one GAD65-specific T cell clone of many identified can transfer diabetes. We have generated transgenic mice on the NOD background expressing a T cell receptor (TCR)-specific for peptide epitope 286-300 (p286) of GAD65. These mice have GAD65-specific CD4(+) T cells, as shown by staining with an I-A(g7)(p286) tetramer reagent. Lymphocytes from these TCR transgenic mice proliferate and make interferon gamma, interleukin (IL)-2, tumor necrosis factor (TNF)-alpha, and IL-10 when stimulated in vitro with GAD65 peptide 286-300, yet these TCR transgenic animals do not spontaneously develop diabetes, and insulitis is virtually undetectable. Furthermore, in vitro activated CD4 T cells from GAD 286 TCR transgenic mice express higher levels of CTL-associated antigen (CTLA)-4 than nontransgenic littermates. CD4(+) T cells, or p286-tetramer(+)CD4(+) Tcells, from GAD65 286-300-specific TCR transgenic mice delay diabetes induced in NOD.scid mice by diabetic NOD spleen cells. This data suggests that GAD65 peptide 286-300-specific T cells have disease protective capacity and are not pathogenic.

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p286/I-Ag7 tetramer-positive CD4+ T cells from G286 mice delay diabetes transfer. Tetramer-positive and -negative cells were sorted from G286 lymph node cells which were activated with irradiated NOD spleen and p286 for 48 h. B220 and CD8 cells were depleted, and the remaining cells were stained with p286 tetramer and CD4 (C, left panel). Gates were set as indicated (C, middle panel), and cells transferred had purity of 98% for sorted populations. The indicated populations were transferred to NOD.scid mice in two separate experiments (A and B). Group sizes are as follows: In A, diabetic spleen cells (n = 10), G286 CD4+ (n = 10), tetramer-high (n = 4), tetramer-negative (n = 4). In B, diabetic spleen cells (n = 15), tetramer-high (n = 8), tetramer-negative (n = 8).
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fig6: p286/I-Ag7 tetramer-positive CD4+ T cells from G286 mice delay diabetes transfer. Tetramer-positive and -negative cells were sorted from G286 lymph node cells which were activated with irradiated NOD spleen and p286 for 48 h. B220 and CD8 cells were depleted, and the remaining cells were stained with p286 tetramer and CD4 (C, left panel). Gates were set as indicated (C, middle panel), and cells transferred had purity of 98% for sorted populations. The indicated populations were transferred to NOD.scid mice in two separate experiments (A and B). Group sizes are as follows: In A, diabetic spleen cells (n = 10), G286 CD4+ (n = 10), tetramer-high (n = 4), tetramer-negative (n = 4). In B, diabetic spleen cells (n = 15), tetramer-high (n = 8), tetramer-negative (n = 8).

Mentions: Statistical significance of delay in diabetes transfer was determined at each time point by Fisher's exact test and is indicated on the incidence plots. All comparisons are between the experimental condition indicated and the control condition consisting of spleen cells from diabetic NOD females transferred alone, except in Fig. 6 A where samples containing tetramer positive cells are compared with those containing tetramer-negative cells. P values < 0.05 are indicated by the symbol *, whereas P values <0.01 are indicated by the symbol #.


CD4(+) T cells from glutamic acid decarboxylase (GAD)65-specific T cell receptor transgenic mice are not diabetogenic and can delay diabetes transfer.

Tarbell KV, Lee M, Ranheim E, Chao CC, Sanna M, Kim SK, Dickie P, Teyton L, Davis M, McDevitt H - J. Exp. Med. (2002)

p286/I-Ag7 tetramer-positive CD4+ T cells from G286 mice delay diabetes transfer. Tetramer-positive and -negative cells were sorted from G286 lymph node cells which were activated with irradiated NOD spleen and p286 for 48 h. B220 and CD8 cells were depleted, and the remaining cells were stained with p286 tetramer and CD4 (C, left panel). Gates were set as indicated (C, middle panel), and cells transferred had purity of 98% for sorted populations. The indicated populations were transferred to NOD.scid mice in two separate experiments (A and B). Group sizes are as follows: In A, diabetic spleen cells (n = 10), G286 CD4+ (n = 10), tetramer-high (n = 4), tetramer-negative (n = 4). In B, diabetic spleen cells (n = 15), tetramer-high (n = 8), tetramer-negative (n = 8).
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196059&req=5

fig6: p286/I-Ag7 tetramer-positive CD4+ T cells from G286 mice delay diabetes transfer. Tetramer-positive and -negative cells were sorted from G286 lymph node cells which were activated with irradiated NOD spleen and p286 for 48 h. B220 and CD8 cells were depleted, and the remaining cells were stained with p286 tetramer and CD4 (C, left panel). Gates were set as indicated (C, middle panel), and cells transferred had purity of 98% for sorted populations. The indicated populations were transferred to NOD.scid mice in two separate experiments (A and B). Group sizes are as follows: In A, diabetic spleen cells (n = 10), G286 CD4+ (n = 10), tetramer-high (n = 4), tetramer-negative (n = 4). In B, diabetic spleen cells (n = 15), tetramer-high (n = 8), tetramer-negative (n = 8).
Mentions: Statistical significance of delay in diabetes transfer was determined at each time point by Fisher's exact test and is indicated on the incidence plots. All comparisons are between the experimental condition indicated and the control condition consisting of spleen cells from diabetic NOD females transferred alone, except in Fig. 6 A where samples containing tetramer positive cells are compared with those containing tetramer-negative cells. P values < 0.05 are indicated by the symbol *, whereas P values <0.01 are indicated by the symbol #.

Bottom Line: However, the exact role of GAD65-specific T cells in diabetes pathogenesis is unclear.Lymphocytes from these TCR transgenic mice proliferate and make interferon gamma, interleukin (IL)-2, tumor necrosis factor (TNF)-alpha, and IL-10 when stimulated in vitro with GAD65 peptide 286-300, yet these TCR transgenic animals do not spontaneously develop diabetes, and insulitis is virtually undetectable.This data suggests that GAD65 peptide 286-300-specific T cells have disease protective capacity and are not pathogenic.

View Article: PubMed Central - PubMed

Affiliation: Program in Immunology, Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA.

ABSTRACT
Glutamic acid decarboxylase (GAD)65 is an early and important antigen in both human diabetes mellitus and the nonobese diabetic (NOD) mouse. However, the exact role of GAD65-specific T cells in diabetes pathogenesis is unclear. T cell responses to GAD65 occur early in diabetes pathogenesis, yet only one GAD65-specific T cell clone of many identified can transfer diabetes. We have generated transgenic mice on the NOD background expressing a T cell receptor (TCR)-specific for peptide epitope 286-300 (p286) of GAD65. These mice have GAD65-specific CD4(+) T cells, as shown by staining with an I-A(g7)(p286) tetramer reagent. Lymphocytes from these TCR transgenic mice proliferate and make interferon gamma, interleukin (IL)-2, tumor necrosis factor (TNF)-alpha, and IL-10 when stimulated in vitro with GAD65 peptide 286-300, yet these TCR transgenic animals do not spontaneously develop diabetes, and insulitis is virtually undetectable. Furthermore, in vitro activated CD4 T cells from GAD 286 TCR transgenic mice express higher levels of CTL-associated antigen (CTLA)-4 than nontransgenic littermates. CD4(+) T cells, or p286-tetramer(+)CD4(+) Tcells, from GAD65 286-300-specific TCR transgenic mice delay diabetes induced in NOD.scid mice by diabetic NOD spleen cells. This data suggests that GAD65 peptide 286-300-specific T cells have disease protective capacity and are not pathogenic.

Show MeSH
Related in: MedlinePlus