Abstract
Cross-linking of glycosylphosphatidylinositol-anchored proteins, including mouse Ly-6A/E, leads to IL-2 secretion and T cell activation, whereas engagement of Ly-6A/E uniquely inhibits IL-2 production induced via TCR. However, little is known concerning the molecular mechanism by which glycosylphosphatidylinositol-anchored proteins regulate IL-2 expression. In this study, we have examined the ability of an anti-Ly-6A/E mAb to regulate transcription factors controlling IL-2 expression. Stimulation of EL4J(Ly-6E).A4 cells with anti-CD3ε or anti-Ly6A/E mAbs induced nuclear factor (NF)-κB p65-p50 (RelA/p50) and AP-1 (Fos/Jun) binding activities and increased nuclear factor of activated T cells (NF-AT) activity, whereas octamer-binding factor and NF-Y levels were stable. Cyclic AMP response element binding protein and T cell-specific factor-1(α) activities were selectively enhanced by anti-CD3ε, but not by anti-Ly6A/E, which suggests that signaling via the TCR and Ly-6 were not identical. Costimulation of these cells with both mAbs produced substantially reduced levels of AP-1, NF-AT, and, especially, NF-κB p65-p50 whereas cyclic AMP response element binding protein and T cell-specific factor-1(α) were induced to a level seen after stimulation by anti-CD3ε. The inducibility of the IL-2 enhancer in vivo and the contribution of individual transcription factors for this induction were assessed with use of reporter chloramphenicol acetyltransferase constructs containing the IL-2 enhancer or oligomerized binding sites for transcription factors. These experiments also demonstrated a key role for NF-κB and AP-1 in the transcriptional regulation of the IL-2 gene by TCR- and Ly6A/E-mediated signaling. By using the 2B4.11 T cell hybridoma and a mutated variant, we revealed a crucial role for the ζ-chain in Ly6A/E-mediated activation of NF-κB.
| Original language | English |
|---|---|
| Pages (from-to) | 2394-2406 |
| Number of pages | 13 |
| Journal | Journal of Immunology |
| Volume | 153 |
| Issue number | 6 |
| State | Published - Sep 15 1994 |
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- Immunology
Cite this
Regulation of nuclear factor-κB and activator protein-1 activities after stimulation of T cells via glycosylphosphatidylinositol-anchored Ly-GA/E. / Ivanov, Vladimir; Fleming, Tony J.; Malek, Thomas.
In: Journal of Immunology, Vol. 153, No. 6, 15.09.1994, p. 2394-2406.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Regulation of nuclear factor-κB and activator protein-1 activities after stimulation of T cells via glycosylphosphatidylinositol-anchored Ly-GA/E
AU - Ivanov, Vladimir
AU - Fleming, Tony J.
AU - Malek, Thomas
PY - 1994/9/15
Y1 - 1994/9/15
N2 - Cross-linking of glycosylphosphatidylinositol-anchored proteins, including mouse Ly-6A/E, leads to IL-2 secretion and T cell activation, whereas engagement of Ly-6A/E uniquely inhibits IL-2 production induced via TCR. However, little is known concerning the molecular mechanism by which glycosylphosphatidylinositol-anchored proteins regulate IL-2 expression. In this study, we have examined the ability of an anti-Ly-6A/E mAb to regulate transcription factors controlling IL-2 expression. Stimulation of EL4J(Ly-6E).A4 cells with anti-CD3ε or anti-Ly6A/E mAbs induced nuclear factor (NF)-κB p65-p50 (RelA/p50) and AP-1 (Fos/Jun) binding activities and increased nuclear factor of activated T cells (NF-AT) activity, whereas octamer-binding factor and NF-Y levels were stable. Cyclic AMP response element binding protein and T cell-specific factor-1(α) activities were selectively enhanced by anti-CD3ε, but not by anti-Ly6A/E, which suggests that signaling via the TCR and Ly-6 were not identical. Costimulation of these cells with both mAbs produced substantially reduced levels of AP-1, NF-AT, and, especially, NF-κB p65-p50 whereas cyclic AMP response element binding protein and T cell-specific factor-1(α) were induced to a level seen after stimulation by anti-CD3ε. The inducibility of the IL-2 enhancer in vivo and the contribution of individual transcription factors for this induction were assessed with use of reporter chloramphenicol acetyltransferase constructs containing the IL-2 enhancer or oligomerized binding sites for transcription factors. These experiments also demonstrated a key role for NF-κB and AP-1 in the transcriptional regulation of the IL-2 gene by TCR- and Ly6A/E-mediated signaling. By using the 2B4.11 T cell hybridoma and a mutated variant, we revealed a crucial role for the ζ-chain in Ly6A/E-mediated activation of NF-κB.
AB - Cross-linking of glycosylphosphatidylinositol-anchored proteins, including mouse Ly-6A/E, leads to IL-2 secretion and T cell activation, whereas engagement of Ly-6A/E uniquely inhibits IL-2 production induced via TCR. However, little is known concerning the molecular mechanism by which glycosylphosphatidylinositol-anchored proteins regulate IL-2 expression. In this study, we have examined the ability of an anti-Ly-6A/E mAb to regulate transcription factors controlling IL-2 expression. Stimulation of EL4J(Ly-6E).A4 cells with anti-CD3ε or anti-Ly6A/E mAbs induced nuclear factor (NF)-κB p65-p50 (RelA/p50) and AP-1 (Fos/Jun) binding activities and increased nuclear factor of activated T cells (NF-AT) activity, whereas octamer-binding factor and NF-Y levels were stable. Cyclic AMP response element binding protein and T cell-specific factor-1(α) activities were selectively enhanced by anti-CD3ε, but not by anti-Ly6A/E, which suggests that signaling via the TCR and Ly-6 were not identical. Costimulation of these cells with both mAbs produced substantially reduced levels of AP-1, NF-AT, and, especially, NF-κB p65-p50 whereas cyclic AMP response element binding protein and T cell-specific factor-1(α) were induced to a level seen after stimulation by anti-CD3ε. The inducibility of the IL-2 enhancer in vivo and the contribution of individual transcription factors for this induction were assessed with use of reporter chloramphenicol acetyltransferase constructs containing the IL-2 enhancer or oligomerized binding sites for transcription factors. These experiments also demonstrated a key role for NF-κB and AP-1 in the transcriptional regulation of the IL-2 gene by TCR- and Ly6A/E-mediated signaling. By using the 2B4.11 T cell hybridoma and a mutated variant, we revealed a crucial role for the ζ-chain in Ly6A/E-mediated activation of NF-κB.
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UR - http://www.scopus.com/inward/citedby.url?scp=0028169802&partnerID=8YFLogxK
M3 - Article
C2 - 7915738
AN - SCOPUS:0028169802
VL - 153
SP - 2394
EP - 2406
JO - Journal of Immunology
JF - Journal of Immunology
SN - 0022-1767
IS - 6
ER -