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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: 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.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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Potential mechanisms by which G286 T cells delay diabetes transfer. (A) CTLA-4 expression on CD4+ cells activated with anti-CD3 and anti-CD28. Results shown were gated on CD4+. The top panel shows CTLA-4 expression for nontransgenic (gray) and G286 (white). Similar results were found in three separate experiments. The bottom panel shows CD4+ cells from 286 mice gated on tetramer-negative (gray) or -positive (white). (B) The role of CD25+ cells in diabetes transfer. The G286 spleen cell population was incubated with biotinylated anti-CD25 and these cells were depleted using streptavidin magnetic beads (Dynal). Diabetic spleen (n = 13; circles); diabetic spleen positive 286 spleen (n = 12; squares); diabetic spleen plus CD25-negative 286 spleen (n = 13; diamonds); CD25-negative 286 spleen (n = 12; triangles).
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fig7: Potential mechanisms by which G286 T cells delay diabetes transfer. (A) CTLA-4 expression on CD4+ cells activated with anti-CD3 and anti-CD28. Results shown were gated on CD4+. The top panel shows CTLA-4 expression for nontransgenic (gray) and G286 (white). Similar results were found in three separate experiments. The bottom panel shows CD4+ cells from 286 mice gated on tetramer-negative (gray) or -positive (white). (B) The role of CD25+ cells in diabetes transfer. The G286 spleen cell population was incubated with biotinylated anti-CD25 and these cells were depleted using streptavidin magnetic beads (Dynal). Diabetic spleen (n = 13; circles); diabetic spleen positive 286 spleen (n = 12; squares); diabetic spleen plus CD25-negative 286 spleen (n = 13; diamonds); CD25-negative 286 spleen (n = 12; triangles).

Mentions: Expression of the regulatory molecule CTLA-4 is one possible mechanism by which CD4+ T cells from G286 mice could mediate protection from diabetes transfer. For example, increased CTLA-4 expression could lead to downmodulation of other pathogenic cells by inducing TGF-β expression (45, 46). When T cells were activated with anti-CD3 and anti–CD28-coated plates, CD4+ lymph node cells from G286 mice expressed higher levels of CTLA-4 than nontransgenic littermates. In addition, when CD4+ T cells from transgenic mice were divided into tetramer positive and tetramer negative subsets, the tetramer positive subset expressed higher levels of CTLA-4 (Fig. 7 A). For the plots shown, the mean fluorescence intensity (MFI) of CTLA-4 expression was 4.3 for CD4+ cells from the transgene negative mice. For G286 mice, the MFI of CD4+ tetramer-negative cells was 5.8, and for CD4+ tetramer-positive cells was 9.0. Furthermore, cells expressing higher levels of tetramer staining had higher MFI for CTLA-4 expression; for the 1% of cells with the highest tetramer staining, the MFI was 12. The ranges of these increases in CTLA-4 expression are similar to those seen when comparing expression on CD4+CD25− versus CD4+CD25+ cells (47).


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)

Potential mechanisms by which G286 T cells delay diabetes transfer. (A) CTLA-4 expression on CD4+ cells activated with anti-CD3 and anti-CD28. Results shown were gated on CD4+. The top panel shows CTLA-4 expression for nontransgenic (gray) and G286 (white). Similar results were found in three separate experiments. The bottom panel shows CD4+ cells from 286 mice gated on tetramer-negative (gray) or -positive (white). (B) The role of CD25+ cells in diabetes transfer. The G286 spleen cell population was incubated with biotinylated anti-CD25 and these cells were depleted using streptavidin magnetic beads (Dynal). Diabetic spleen (n = 13; circles); diabetic spleen positive 286 spleen (n = 12; squares); diabetic spleen plus CD25-negative 286 spleen (n = 13; diamonds); CD25-negative 286 spleen (n = 12; triangles).
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Related In: Results  -  Collection

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

fig7: Potential mechanisms by which G286 T cells delay diabetes transfer. (A) CTLA-4 expression on CD4+ cells activated with anti-CD3 and anti-CD28. Results shown were gated on CD4+. The top panel shows CTLA-4 expression for nontransgenic (gray) and G286 (white). Similar results were found in three separate experiments. The bottom panel shows CD4+ cells from 286 mice gated on tetramer-negative (gray) or -positive (white). (B) The role of CD25+ cells in diabetes transfer. The G286 spleen cell population was incubated with biotinylated anti-CD25 and these cells were depleted using streptavidin magnetic beads (Dynal). Diabetic spleen (n = 13; circles); diabetic spleen positive 286 spleen (n = 12; squares); diabetic spleen plus CD25-negative 286 spleen (n = 13; diamonds); CD25-negative 286 spleen (n = 12; triangles).
Mentions: Expression of the regulatory molecule CTLA-4 is one possible mechanism by which CD4+ T cells from G286 mice could mediate protection from diabetes transfer. For example, increased CTLA-4 expression could lead to downmodulation of other pathogenic cells by inducing TGF-β expression (45, 46). When T cells were activated with anti-CD3 and anti–CD28-coated plates, CD4+ lymph node cells from G286 mice expressed higher levels of CTLA-4 than nontransgenic littermates. In addition, when CD4+ T cells from transgenic mice were divided into tetramer positive and tetramer negative subsets, the tetramer positive subset expressed higher levels of CTLA-4 (Fig. 7 A). For the plots shown, the mean fluorescence intensity (MFI) of CTLA-4 expression was 4.3 for CD4+ cells from the transgene negative mice. For G286 mice, the MFI of CD4+ tetramer-negative cells was 5.8, and for CD4+ tetramer-positive cells was 9.0. Furthermore, cells expressing higher levels of tetramer staining had higher MFI for CTLA-4 expression; for the 1% of cells with the highest tetramer staining, the MFI was 12. The ranges of these increases in CTLA-4 expression are similar to those seen when comparing expression on CD4+CD25− versus CD4+CD25+ cells (47).

Bottom Line: 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.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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Related in: MedlinePlus