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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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Evidence for expression of multiple TCRs in G286 mice. (A) Peripheral blood lymphocytes from G286 Cα−/− (left) G286 Cα+/− (middle) and NOD Cα−/− (right) mice were stained with antibodies for CD4 and Vβ8 (top) or CD8 and Vα2 (bottom). The percentage of cells in each quadrant are indicated in each corner of the plots. (B) Cells from lymph node of a G286 mouse or thymus of a G286 Cα+/− mouse were stained with antibodies for CD4, CD8, and CD3. Cells were gated on CD4+ or CD8+. Expression of CD3 versus I-Ag7-286 tetramer is shown. Cells in the diagonal gate represent cells for which the p286-specific TCR is the predominant TCR expressed. Percentage of cells in the gates drawn are indicated on the plots.
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fig2: Evidence for expression of multiple TCRs in G286 mice. (A) Peripheral blood lymphocytes from G286 Cα−/− (left) G286 Cα+/− (middle) and NOD Cα−/− (right) mice were stained with antibodies for CD4 and Vβ8 (top) or CD8 and Vα2 (bottom). The percentage of cells in each quadrant are indicated in each corner of the plots. (B) Cells from lymph node of a G286 mouse or thymus of a G286 Cα+/− mouse were stained with antibodies for CD4, CD8, and CD3. Cells were gated on CD4+ or CD8+. Expression of CD3 versus I-Ag7-286 tetramer is shown. Cells in the diagonal gate represent cells for which the p286-specific TCR is the predominant TCR expressed. Percentage of cells in the gates drawn are indicated on the plots.

Mentions: G286 mice were crossed onto NOD TCR Cα−/− mice to determine the role of T cells expressing endogenous TCR-α chain in selection and disease. In MHC class II–restricted TCR transgenics crossed onto Cα−/−, one would expect CD4+ cells to increase and CD8+ cells to decrease due to a decrease in the endogenous repertoire. Surprisingly, fewer CD4+ T cells were found in the periphery, and the percentage of CD8+ cells was not diminished in G286 Cα−/− mice. The lack of Vα2+ cells confirms that these mice lack expression of endogenous α chains and are TCR-Cα−/−. Many (20%) of the CD4+ cells from G286 Cα−/− mice express Vβ8, indicating that allelic exclusion of Vβ is not complete (Fig. 2 A). The most likely explanation for this result is that endogenous TCR-α chain expression in G286 Cα+/+ NOD mice allows this autoreactive T cell to survive thymic negative selection; the few CD4+ cells in G286 Cα−/− NOD mice are compensating by rearranging endogenous Vβ genes. Coexpression of an endogenous TCR-α chain paired with a transgenic TCR-β chain may allow for sufficient downregulation of the transgenic autoreactive TCR to allow escape from thymic negative selection without failure to be positively selected. This phenomenon has been observed with other autoreactive TCRs (37).


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)

Evidence for expression of multiple TCRs in G286 mice. (A) Peripheral blood lymphocytes from G286 Cα−/− (left) G286 Cα+/− (middle) and NOD Cα−/− (right) mice were stained with antibodies for CD4 and Vβ8 (top) or CD8 and Vα2 (bottom). The percentage of cells in each quadrant are indicated in each corner of the plots. (B) Cells from lymph node of a G286 mouse or thymus of a G286 Cα+/− mouse were stained with antibodies for CD4, CD8, and CD3. Cells were gated on CD4+ or CD8+. Expression of CD3 versus I-Ag7-286 tetramer is shown. Cells in the diagonal gate represent cells for which the p286-specific TCR is the predominant TCR expressed. Percentage of cells in the gates drawn are indicated on the plots.
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Related In: Results  -  Collection

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

fig2: Evidence for expression of multiple TCRs in G286 mice. (A) Peripheral blood lymphocytes from G286 Cα−/− (left) G286 Cα+/− (middle) and NOD Cα−/− (right) mice were stained with antibodies for CD4 and Vβ8 (top) or CD8 and Vα2 (bottom). The percentage of cells in each quadrant are indicated in each corner of the plots. (B) Cells from lymph node of a G286 mouse or thymus of a G286 Cα+/− mouse were stained with antibodies for CD4, CD8, and CD3. Cells were gated on CD4+ or CD8+. Expression of CD3 versus I-Ag7-286 tetramer is shown. Cells in the diagonal gate represent cells for which the p286-specific TCR is the predominant TCR expressed. Percentage of cells in the gates drawn are indicated on the plots.
Mentions: G286 mice were crossed onto NOD TCR Cα−/− mice to determine the role of T cells expressing endogenous TCR-α chain in selection and disease. In MHC class II–restricted TCR transgenics crossed onto Cα−/−, one would expect CD4+ cells to increase and CD8+ cells to decrease due to a decrease in the endogenous repertoire. Surprisingly, fewer CD4+ T cells were found in the periphery, and the percentage of CD8+ cells was not diminished in G286 Cα−/− mice. The lack of Vα2+ cells confirms that these mice lack expression of endogenous α chains and are TCR-Cα−/−. Many (20%) of the CD4+ cells from G286 Cα−/− mice express Vβ8, indicating that allelic exclusion of Vβ is not complete (Fig. 2 A). The most likely explanation for this result is that endogenous TCR-α chain expression in G286 Cα+/+ NOD mice allows this autoreactive T cell to survive thymic negative selection; the few CD4+ cells in G286 Cα−/− NOD mice are compensating by rearranging endogenous Vβ genes. Coexpression of an endogenous TCR-α chain paired with a transgenic TCR-β chain may allow for sufficient downregulation of the transgenic autoreactive TCR to allow escape from thymic negative selection without failure to be positively selected. This phenomenon has been observed with other autoreactive TCRs (37).

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