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Gamma delta T cells from tolerized alpha beta T cell receptor (TCR)-deficient mice inhibit contact sensitivity-effector T cells in vivo, and their interferon-gamma production in vitro.

Szczepanik M, Anderson LR, Ushio H, Ptak W, Owen MJ, Hayday AC, Askenase PW - J. Exp. Med. (1996)

Bottom Line: Downregulation by gamma delta cells showed specificity for hapten, but was not restricted by the MHC.Together, these findings establish that gamma delta T cells cannot fulfill CS-effector functions performed by alpha beta T cells, but may fulfill an Ag-specific downregulatory role that may be directly comparable to reports of Ag-specific downregulation of IgE antibody responses by gamma delta T cells.Comparisons are likewise considered with downregulation by gamma delta T cells occurring in immune responses to pathogens, tumors, and allografts, and in systemic autoimmunity.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, College of Medicine, Jagiellonian University, Krakow, Poland.

ABSTRACT
Contact sensitivity (CS) responses to reactive hapten Ag, such as picryl chloride (PCl) or oxazolone (OX), are classical examples of T cell-mediated immune responses in vivo that are clearly subject to multifaceted regulation. There is abundant evidence that downregulation of CS may be mediated by T cells exposed to high doses of Ag. This is termed high dose Ag tolerance. To clarify the T cell types that effect CS responses and mediate their downregulation, we have undertaken studies of CS in mice congenitally deficient in specific subsets of lymphocytes. The first such studies, using alpha beta T cell-deficient (TCR alpha -/-) mice, are presented here. The results clearly show that TCR alpha -/- mice cannot mount CS, implicating alpha beta T cells as the critical CS-effector cells. However, TCR alpha -/- mice can, after high dose tolerance, downregulate alpha +/+ CS-effector T cells adoptively transferred into them. By mixing ex vivo and then adoptive cell transfers in vivo, the active downregulatory cells in tolerized alpha -/- mice are shown to include gamma delta TCR+ cells that also can downregulate interferon-gamma production by the targeted CS-effector cells in vitro. Downregulation by gamma delta cells showed specificity for hapten, but was not restricted by the MHC. Together, these findings establish that gamma delta T cells cannot fulfill CS-effector functions performed by alpha beta T cells, but may fulfill an Ag-specific downregulatory role that may be directly comparable to reports of Ag-specific downregulation of IgE antibody responses by gamma delta T cells. Comparisons are likewise considered with downregulation by gamma delta T cells occurring in immune responses to pathogens, tumors, and allografts, and in systemic autoimmunity.

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Lack of MHC restriction of γδ downregulatory cells from  TNBSA-tolerized TCRα−/− mice. Magnetic bead positive (γδ TCR+)  cells from TNBSA-tolerized α−/− mice were incubated with 7 x 107 cells  from MHC compatible 129/J H2b,d, or MHC incompatible (CBA/J,  H2k) mice for 30 min at 37°C (groups B and D). As a positive control, we  used immune cells from 129/J or CBA/J mice that were incubated without regulatory cells in medium alone (groups A and C). Then cells were  washed and injected intraperitoneally into recipients that were syngeneic  with the immune cells, and were skin tested for CS responsiveness. Statistical significance: group A vs B, and group C vs D, P <0.001.
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Figure 5: Lack of MHC restriction of γδ downregulatory cells from TNBSA-tolerized TCRα−/− mice. Magnetic bead positive (γδ TCR+) cells from TNBSA-tolerized α−/− mice were incubated with 7 x 107 cells from MHC compatible 129/J H2b,d, or MHC incompatible (CBA/J, H2k) mice for 30 min at 37°C (groups B and D). As a positive control, we used immune cells from 129/J or CBA/J mice that were incubated without regulatory cells in medium alone (groups A and C). Then cells were washed and injected intraperitoneally into recipients that were syngeneic with the immune cells, and were skin tested for CS responsiveness. Statistical significance: group A vs B, and group C vs D, P <0.001.

Mentions: Spleen cells (1.5 × 108) from four TNBSA-tolerized α+/+ 129/J, or α−/− donor mice were incubated in 20 ml UC7 hamster anti-TCR γδ mAb (12 μg/ml) hybridoma supernatant for 60 min on ice. After mAb coating, cells were washed three times with PBS + 2% FCS, and resuspended in 40 ml PBS + 2% FCS containing goat anti–hamster Ig–coated paramagnetic beads at 5–10 beads per target cell. The cells were then incubated in a flat vertical 50-ml flask for 30 min on ice, after which a magnet (Advanced Magnetics Inc., Cambridge, MA) was applied to one side of the flask. 10 min later, magnetic bead nonadherent cells were recovered, and then the magnetic bead adherent cells were recovered. Nonseparated regulatory cells were used in experiments shown in Figs. 3, 5, and 6 b.


Gamma delta T cells from tolerized alpha beta T cell receptor (TCR)-deficient mice inhibit contact sensitivity-effector T cells in vivo, and their interferon-gamma production in vitro.

Szczepanik M, Anderson LR, Ushio H, Ptak W, Owen MJ, Hayday AC, Askenase PW - J. Exp. Med. (1996)

Lack of MHC restriction of γδ downregulatory cells from  TNBSA-tolerized TCRα−/− mice. Magnetic bead positive (γδ TCR+)  cells from TNBSA-tolerized α−/− mice were incubated with 7 x 107 cells  from MHC compatible 129/J H2b,d, or MHC incompatible (CBA/J,  H2k) mice for 30 min at 37°C (groups B and D). As a positive control, we  used immune cells from 129/J or CBA/J mice that were incubated without regulatory cells in medium alone (groups A and C). Then cells were  washed and injected intraperitoneally into recipients that were syngeneic  with the immune cells, and were skin tested for CS responsiveness. Statistical significance: group A vs B, and group C vs D, P <0.001.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196372&req=5

Figure 5: Lack of MHC restriction of γδ downregulatory cells from TNBSA-tolerized TCRα−/− mice. Magnetic bead positive (γδ TCR+) cells from TNBSA-tolerized α−/− mice were incubated with 7 x 107 cells from MHC compatible 129/J H2b,d, or MHC incompatible (CBA/J, H2k) mice for 30 min at 37°C (groups B and D). As a positive control, we used immune cells from 129/J or CBA/J mice that were incubated without regulatory cells in medium alone (groups A and C). Then cells were washed and injected intraperitoneally into recipients that were syngeneic with the immune cells, and were skin tested for CS responsiveness. Statistical significance: group A vs B, and group C vs D, P <0.001.
Mentions: Spleen cells (1.5 × 108) from four TNBSA-tolerized α+/+ 129/J, or α−/− donor mice were incubated in 20 ml UC7 hamster anti-TCR γδ mAb (12 μg/ml) hybridoma supernatant for 60 min on ice. After mAb coating, cells were washed three times with PBS + 2% FCS, and resuspended in 40 ml PBS + 2% FCS containing goat anti–hamster Ig–coated paramagnetic beads at 5–10 beads per target cell. The cells were then incubated in a flat vertical 50-ml flask for 30 min on ice, after which a magnet (Advanced Magnetics Inc., Cambridge, MA) was applied to one side of the flask. 10 min later, magnetic bead nonadherent cells were recovered, and then the magnetic bead adherent cells were recovered. Nonseparated regulatory cells were used in experiments shown in Figs. 3, 5, and 6 b.

Bottom Line: Downregulation by gamma delta cells showed specificity for hapten, but was not restricted by the MHC.Together, these findings establish that gamma delta T cells cannot fulfill CS-effector functions performed by alpha beta T cells, but may fulfill an Ag-specific downregulatory role that may be directly comparable to reports of Ag-specific downregulation of IgE antibody responses by gamma delta T cells.Comparisons are likewise considered with downregulation by gamma delta T cells occurring in immune responses to pathogens, tumors, and allografts, and in systemic autoimmunity.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, College of Medicine, Jagiellonian University, Krakow, Poland.

ABSTRACT
Contact sensitivity (CS) responses to reactive hapten Ag, such as picryl chloride (PCl) or oxazolone (OX), are classical examples of T cell-mediated immune responses in vivo that are clearly subject to multifaceted regulation. There is abundant evidence that downregulation of CS may be mediated by T cells exposed to high doses of Ag. This is termed high dose Ag tolerance. To clarify the T cell types that effect CS responses and mediate their downregulation, we have undertaken studies of CS in mice congenitally deficient in specific subsets of lymphocytes. The first such studies, using alpha beta T cell-deficient (TCR alpha -/-) mice, are presented here. The results clearly show that TCR alpha -/- mice cannot mount CS, implicating alpha beta T cells as the critical CS-effector cells. However, TCR alpha -/- mice can, after high dose tolerance, downregulate alpha +/+ CS-effector T cells adoptively transferred into them. By mixing ex vivo and then adoptive cell transfers in vivo, the active downregulatory cells in tolerized alpha -/- mice are shown to include gamma delta TCR+ cells that also can downregulate interferon-gamma production by the targeted CS-effector cells in vitro. Downregulation by gamma delta cells showed specificity for hapten, but was not restricted by the MHC. Together, these findings establish that gamma delta T cells cannot fulfill CS-effector functions performed by alpha beta T cells, but may fulfill an Ag-specific downregulatory role that may be directly comparable to reports of Ag-specific downregulation of IgE antibody responses by gamma delta T cells. Comparisons are likewise considered with downregulation by gamma delta T cells occurring in immune responses to pathogens, tumors, and allografts, and in systemic autoimmunity.

Show MeSH
Related in: MedlinePlus