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Neonatal tumor necrosis factor alpha promotes diabetes in nonobese diabetic mice by CD154-independent antigen presentation to CD8(+) T cells.

Green EA, Wong FS, Eshima K, Mora C, Flavell RA - J. Exp. Med. (2000)

Bottom Line: Here we show that TNF-alpha promotes autoaggression of both effector CD4(+) and CD8(+) T cells.Instead, it appears that TNF-alpha can promote cross-presentation of islet antigen to CD8(+) T cells using a unique CD40-CD154-independent pathway.These data provide new insights into the mechanisms by which inflammatory stimuli can bypass CD154-CD40 immune regulatory signals and cause activation of autoreactive T cells.

View Article: PubMed Central - PubMed

Affiliation: Section of Immunobiology, Yale University School of Medicine, New Haven, Connecticut 06520, USA.

ABSTRACT
Neonatal islet-specific expression of tumor necrosis factor (TNF)-alpha in nonobese diabetic mice promotes diabetes by provoking islet-infiltrating antigen-presenting cells to present islet peptides to autoreactive T cells. Here we show that TNF-alpha promotes autoaggression of both effector CD4(+) and CD8(+) T cells. Whereas CD8(+) T cells are critical for diabetes progression, CD4(+) T cells play a lesser role. TNF-alpha-mediated diabetes development was not dependent on CD154-CD40 signals or activated CD4(+) T cells. Instead, it appears that TNF-alpha can promote cross-presentation of islet antigen to CD8(+) T cells using a unique CD40-CD154-independent pathway. These data provide new insights into the mechanisms by which inflammatory stimuli can bypass CD154-CD40 immune regulatory signals and cause activation of autoreactive T cells.

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TNF-α–CD154−/− mice cannot mount GAD-specific CD4+ T cell responses. (a) CD4+ T cells were purified from the pancreatic lymph nodes of 6-wk-old TNF-α–CD154+/+ (♦), TNF-α–CD154−/− (○), or non-tg NOD–CD154−/− (▴) mice and cultured with 5 × 105 irradiated splenocytes from 5-wk-old NOD mice and 0–20 μg of GADp35. After a 4-d culture, proliferative responses of triplicate cultures were determined. Background cpm for CD4+ T cells cultured with irradiated splenocytes was <500. (b) CD4+ T cells from TNF-α–CD154−/− mice show decreased responses to islet antigen. Left, irradiated islet-infiltrating cells (3 × 104) from 6-wk-old TNF-α–CD154+/+ or TNF-α–CD154−/− mice were cultured with 2 × 106 CD4+ T cells purified from 6-wk-old TNF-α–CD154−/− mice or age-matched control NOD mice. Proliferative responses of triplicate cultures were measured after 4 d as described previously. Background cpm for CD4+ T cells was <520. Right, islet-infiltrating cells from 6-wk-old TNF-α–CD154+/+ or TNF-α–CD154−/− mice were depleted of CD8+ T cells, and the remaining cells were cultured with 2.5 μg/ml of Con A for 48 h. Culture supernatants were harvested at the end of the incubation period, and IFN-γ was detected by ELISA. The ELISA assay was performed in triplicate and the data presented as mean ± SD. (c) CD4+ T cells in TNF-α–CD154−/− mice cannot be primed to exogenous protein antigens. Groups of two 6-wk-old TNF-α–CD154+/+ (♦), TNF-α–CD154−/− (▴), non-tg NOD–CD154+/+ (▪), and non-tg NOD–CD154−/− (•) mice were immunized subcutaneously via their footpads with 100 μg of KLH in alum. 7 d later, 5 × 105 cells isolated from the popliteal lymph nodes were cultured with KLH at the concentrations (conc.) shown. After a 4-d incubation, proliferative responses of duplicate cultures were measured as before. Background cpm values for cells cultured in medium alone have been subtracted from the data.
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Figure 4: TNF-α–CD154−/− mice cannot mount GAD-specific CD4+ T cell responses. (a) CD4+ T cells were purified from the pancreatic lymph nodes of 6-wk-old TNF-α–CD154+/+ (♦), TNF-α–CD154−/− (○), or non-tg NOD–CD154−/− (▴) mice and cultured with 5 × 105 irradiated splenocytes from 5-wk-old NOD mice and 0–20 μg of GADp35. After a 4-d culture, proliferative responses of triplicate cultures were determined. Background cpm for CD4+ T cells cultured with irradiated splenocytes was <500. (b) CD4+ T cells from TNF-α–CD154−/− mice show decreased responses to islet antigen. Left, irradiated islet-infiltrating cells (3 × 104) from 6-wk-old TNF-α–CD154+/+ or TNF-α–CD154−/− mice were cultured with 2 × 106 CD4+ T cells purified from 6-wk-old TNF-α–CD154−/− mice or age-matched control NOD mice. Proliferative responses of triplicate cultures were measured after 4 d as described previously. Background cpm for CD4+ T cells was <520. Right, islet-infiltrating cells from 6-wk-old TNF-α–CD154+/+ or TNF-α–CD154−/− mice were depleted of CD8+ T cells, and the remaining cells were cultured with 2.5 μg/ml of Con A for 48 h. Culture supernatants were harvested at the end of the incubation period, and IFN-γ was detected by ELISA. The ELISA assay was performed in triplicate and the data presented as mean ± SD. (c) CD4+ T cells in TNF-α–CD154−/− mice cannot be primed to exogenous protein antigens. Groups of two 6-wk-old TNF-α–CD154+/+ (♦), TNF-α–CD154−/− (▴), non-tg NOD–CD154+/+ (▪), and non-tg NOD–CD154−/− (•) mice were immunized subcutaneously via their footpads with 100 μg of KLH in alum. 7 d later, 5 × 105 cells isolated from the popliteal lymph nodes were cultured with KLH at the concentrations (conc.) shown. After a 4-d incubation, proliferative responses of duplicate cultures were measured as before. Background cpm values for cells cultured in medium alone have been subtracted from the data.

Mentions: Neither CD4+ T cells from TNF-α–CD154−/− mice nor non-tg NOD–CD154−/− mice responded to GADp35, which contrasted significantly to the vigorous response by CD4+ T cells from TNF-α–CD154+/+ mice (Fig. 4 a). To confirm that this lack of response in the latter two groups of mice did not relate to the GAD peptide used, we repeated the experiments with whole GAD protein. Similar to the above findings, only CD4+ T cells from TNF-α–CD154+/+ mice responded to GAD (not shown).


Neonatal tumor necrosis factor alpha promotes diabetes in nonobese diabetic mice by CD154-independent antigen presentation to CD8(+) T cells.

Green EA, Wong FS, Eshima K, Mora C, Flavell RA - J. Exp. Med. (2000)

TNF-α–CD154−/− mice cannot mount GAD-specific CD4+ T cell responses. (a) CD4+ T cells were purified from the pancreatic lymph nodes of 6-wk-old TNF-α–CD154+/+ (♦), TNF-α–CD154−/− (○), or non-tg NOD–CD154−/− (▴) mice and cultured with 5 × 105 irradiated splenocytes from 5-wk-old NOD mice and 0–20 μg of GADp35. After a 4-d culture, proliferative responses of triplicate cultures were determined. Background cpm for CD4+ T cells cultured with irradiated splenocytes was <500. (b) CD4+ T cells from TNF-α–CD154−/− mice show decreased responses to islet antigen. Left, irradiated islet-infiltrating cells (3 × 104) from 6-wk-old TNF-α–CD154+/+ or TNF-α–CD154−/− mice were cultured with 2 × 106 CD4+ T cells purified from 6-wk-old TNF-α–CD154−/− mice or age-matched control NOD mice. Proliferative responses of triplicate cultures were measured after 4 d as described previously. Background cpm for CD4+ T cells was <520. Right, islet-infiltrating cells from 6-wk-old TNF-α–CD154+/+ or TNF-α–CD154−/− mice were depleted of CD8+ T cells, and the remaining cells were cultured with 2.5 μg/ml of Con A for 48 h. Culture supernatants were harvested at the end of the incubation period, and IFN-γ was detected by ELISA. The ELISA assay was performed in triplicate and the data presented as mean ± SD. (c) CD4+ T cells in TNF-α–CD154−/− mice cannot be primed to exogenous protein antigens. Groups of two 6-wk-old TNF-α–CD154+/+ (♦), TNF-α–CD154−/− (▴), non-tg NOD–CD154+/+ (▪), and non-tg NOD–CD154−/− (•) mice were immunized subcutaneously via their footpads with 100 μg of KLH in alum. 7 d later, 5 × 105 cells isolated from the popliteal lymph nodes were cultured with KLH at the concentrations (conc.) shown. After a 4-d incubation, proliferative responses of duplicate cultures were measured as before. Background cpm values for cells cultured in medium alone have been subtracted from the data.
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Related In: Results  -  Collection

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Figure 4: TNF-α–CD154−/− mice cannot mount GAD-specific CD4+ T cell responses. (a) CD4+ T cells were purified from the pancreatic lymph nodes of 6-wk-old TNF-α–CD154+/+ (♦), TNF-α–CD154−/− (○), or non-tg NOD–CD154−/− (▴) mice and cultured with 5 × 105 irradiated splenocytes from 5-wk-old NOD mice and 0–20 μg of GADp35. After a 4-d culture, proliferative responses of triplicate cultures were determined. Background cpm for CD4+ T cells cultured with irradiated splenocytes was <500. (b) CD4+ T cells from TNF-α–CD154−/− mice show decreased responses to islet antigen. Left, irradiated islet-infiltrating cells (3 × 104) from 6-wk-old TNF-α–CD154+/+ or TNF-α–CD154−/− mice were cultured with 2 × 106 CD4+ T cells purified from 6-wk-old TNF-α–CD154−/− mice or age-matched control NOD mice. Proliferative responses of triplicate cultures were measured after 4 d as described previously. Background cpm for CD4+ T cells was <520. Right, islet-infiltrating cells from 6-wk-old TNF-α–CD154+/+ or TNF-α–CD154−/− mice were depleted of CD8+ T cells, and the remaining cells were cultured with 2.5 μg/ml of Con A for 48 h. Culture supernatants were harvested at the end of the incubation period, and IFN-γ was detected by ELISA. The ELISA assay was performed in triplicate and the data presented as mean ± SD. (c) CD4+ T cells in TNF-α–CD154−/− mice cannot be primed to exogenous protein antigens. Groups of two 6-wk-old TNF-α–CD154+/+ (♦), TNF-α–CD154−/− (▴), non-tg NOD–CD154+/+ (▪), and non-tg NOD–CD154−/− (•) mice were immunized subcutaneously via their footpads with 100 μg of KLH in alum. 7 d later, 5 × 105 cells isolated from the popliteal lymph nodes were cultured with KLH at the concentrations (conc.) shown. After a 4-d incubation, proliferative responses of duplicate cultures were measured as before. Background cpm values for cells cultured in medium alone have been subtracted from the data.
Mentions: Neither CD4+ T cells from TNF-α–CD154−/− mice nor non-tg NOD–CD154−/− mice responded to GADp35, which contrasted significantly to the vigorous response by CD4+ T cells from TNF-α–CD154+/+ mice (Fig. 4 a). To confirm that this lack of response in the latter two groups of mice did not relate to the GAD peptide used, we repeated the experiments with whole GAD protein. Similar to the above findings, only CD4+ T cells from TNF-α–CD154+/+ mice responded to GAD (not shown).

Bottom Line: Here we show that TNF-alpha promotes autoaggression of both effector CD4(+) and CD8(+) T cells.Instead, it appears that TNF-alpha can promote cross-presentation of islet antigen to CD8(+) T cells using a unique CD40-CD154-independent pathway.These data provide new insights into the mechanisms by which inflammatory stimuli can bypass CD154-CD40 immune regulatory signals and cause activation of autoreactive T cells.

View Article: PubMed Central - PubMed

Affiliation: Section of Immunobiology, Yale University School of Medicine, New Haven, Connecticut 06520, USA.

ABSTRACT
Neonatal islet-specific expression of tumor necrosis factor (TNF)-alpha in nonobese diabetic mice promotes diabetes by provoking islet-infiltrating antigen-presenting cells to present islet peptides to autoreactive T cells. Here we show that TNF-alpha promotes autoaggression of both effector CD4(+) and CD8(+) T cells. Whereas CD8(+) T cells are critical for diabetes progression, CD4(+) T cells play a lesser role. TNF-alpha-mediated diabetes development was not dependent on CD154-CD40 signals or activated CD4(+) T cells. Instead, it appears that TNF-alpha can promote cross-presentation of islet antigen to CD8(+) T cells using a unique CD40-CD154-independent pathway. These data provide new insights into the mechanisms by which inflammatory stimuli can bypass CD154-CD40 immune regulatory signals and cause activation of autoreactive T cells.

Show MeSH
Related in: MedlinePlus