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Lineage-restricted function of nuclear factor kappaB-inducing kinase (NIK) in transducing signals via CD40.

Garceau N, Kosaka Y, Masters S, Hambor J, Shinkura R, Honjo T, Noelle RJ - J. Exp. Med. (2000)

Bottom Line: Nuclear factor kappaB (NF-kappaB)-inducing kinase (NIK) has been implicated as a central transducing kinase in CD40-dependent activation.Here, we show that although NIK is essential for B cell activation, it is dispensable for activation of DCs.Such data provide compelling evidence that different intermediary kinases are used by different cellular lineages to trigger NF-kappaB activation via CD40.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Dartmouth Medical School, Lebanon, New Hampshire 03756, USA.

ABSTRACT
CD40 signaling in B cells and dendritic cells (DCs) is critical for the development of humoral and cell-mediated immunity, respectively. Nuclear factor kappaB (NF-kappaB)-inducing kinase (NIK) has been implicated as a central transducing kinase in CD40-dependent activation. Here, we show that although NIK is essential for B cell activation, it is dispensable for activation of DCs. Such data provide compelling evidence that different intermediary kinases are used by different cellular lineages to trigger NF-kappaB activation via CD40.

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Role of NIK in activation of B cells. aly/aly and aly/+ splenic B cells were cultured in vitro and assayed for their ability to proliferate (A), produce Ig (B), and upregulate cell surface markers (C) in response to CD40 stimulation. (A) Induction of proliferation by anti-CD40 (10 μg/ml), FGK115, LPS (50 μg/ml), and anti-IgM (goat anti–mouse IgM) was measured by the incorporation of [3H]thymidine from 66 to 72 h after initiation of culture. All cultures contained 10 ng/ml of IL-4. (B) Induction of Ig secretion was performed using sCD154 or LPS in combination with IL-4 (10 ng/ml) and IL-5 (10 ng/ml). Ig secretion was measured using an isotype-specific ELISA. (C) Expression of cell surface molecules on splenic B cells from aly/+ (top) or aly/aly (bottom) mice after culture with (purple histogram) or without (green outline histogram) sCD154 for 48 h was measured by flow cytometry.
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Figure 1: Role of NIK in activation of B cells. aly/aly and aly/+ splenic B cells were cultured in vitro and assayed for their ability to proliferate (A), produce Ig (B), and upregulate cell surface markers (C) in response to CD40 stimulation. (A) Induction of proliferation by anti-CD40 (10 μg/ml), FGK115, LPS (50 μg/ml), and anti-IgM (goat anti–mouse IgM) was measured by the incorporation of [3H]thymidine from 66 to 72 h after initiation of culture. All cultures contained 10 ng/ml of IL-4. (B) Induction of Ig secretion was performed using sCD154 or LPS in combination with IL-4 (10 ng/ml) and IL-5 (10 ng/ml). Ig secretion was measured using an isotype-specific ELISA. (C) Expression of cell surface molecules on splenic B cells from aly/+ (top) or aly/aly (bottom) mice after culture with (purple histogram) or without (green outline histogram) sCD154 for 48 h was measured by flow cytometry.

Mentions: To investigate the function of NIK in CD40-mediated B cell activation, B cells from aly/+ and aly/aly mice were assessed for their ability to proliferate, produce Ig, and upregulate costimulatory molecules in response to soluble (s)CD154 and other polyclonal activators. B cells from aly/aly mice displayed a significant reduction in proliferative capacity in response to CD40, LPS, and anti-Ig activation relative to B cells from aly/+ mice (Fig. 1 A). Similarly, the capacity of B cells from aly/aly mice to produce IgM and IgG in response to sCD154 and LPS was also reduced (Fig. 1 B). In contrast to the diminished proliferation and Ig production observed in the aly/aly B cells, upregulation of several surface molecules (MHC class II, intercellular adhesion molecule 1 [ICAM-1], CD23, and B7.2) that are hallmarks of B cell activation in response to CD40 stimulation appeared intact. However, upregulation of one surface marker, B7.1, was impaired by the aly mutation (Fig. 1 C).


Lineage-restricted function of nuclear factor kappaB-inducing kinase (NIK) in transducing signals via CD40.

Garceau N, Kosaka Y, Masters S, Hambor J, Shinkura R, Honjo T, Noelle RJ - J. Exp. Med. (2000)

Role of NIK in activation of B cells. aly/aly and aly/+ splenic B cells were cultured in vitro and assayed for their ability to proliferate (A), produce Ig (B), and upregulate cell surface markers (C) in response to CD40 stimulation. (A) Induction of proliferation by anti-CD40 (10 μg/ml), FGK115, LPS (50 μg/ml), and anti-IgM (goat anti–mouse IgM) was measured by the incorporation of [3H]thymidine from 66 to 72 h after initiation of culture. All cultures contained 10 ng/ml of IL-4. (B) Induction of Ig secretion was performed using sCD154 or LPS in combination with IL-4 (10 ng/ml) and IL-5 (10 ng/ml). Ig secretion was measured using an isotype-specific ELISA. (C) Expression of cell surface molecules on splenic B cells from aly/+ (top) or aly/aly (bottom) mice after culture with (purple histogram) or without (green outline histogram) sCD154 for 48 h was measured by flow cytometry.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2195761&req=5

Figure 1: Role of NIK in activation of B cells. aly/aly and aly/+ splenic B cells were cultured in vitro and assayed for their ability to proliferate (A), produce Ig (B), and upregulate cell surface markers (C) in response to CD40 stimulation. (A) Induction of proliferation by anti-CD40 (10 μg/ml), FGK115, LPS (50 μg/ml), and anti-IgM (goat anti–mouse IgM) was measured by the incorporation of [3H]thymidine from 66 to 72 h after initiation of culture. All cultures contained 10 ng/ml of IL-4. (B) Induction of Ig secretion was performed using sCD154 or LPS in combination with IL-4 (10 ng/ml) and IL-5 (10 ng/ml). Ig secretion was measured using an isotype-specific ELISA. (C) Expression of cell surface molecules on splenic B cells from aly/+ (top) or aly/aly (bottom) mice after culture with (purple histogram) or without (green outline histogram) sCD154 for 48 h was measured by flow cytometry.
Mentions: To investigate the function of NIK in CD40-mediated B cell activation, B cells from aly/+ and aly/aly mice were assessed for their ability to proliferate, produce Ig, and upregulate costimulatory molecules in response to soluble (s)CD154 and other polyclonal activators. B cells from aly/aly mice displayed a significant reduction in proliferative capacity in response to CD40, LPS, and anti-Ig activation relative to B cells from aly/+ mice (Fig. 1 A). Similarly, the capacity of B cells from aly/aly mice to produce IgM and IgG in response to sCD154 and LPS was also reduced (Fig. 1 B). In contrast to the diminished proliferation and Ig production observed in the aly/aly B cells, upregulation of several surface molecules (MHC class II, intercellular adhesion molecule 1 [ICAM-1], CD23, and B7.2) that are hallmarks of B cell activation in response to CD40 stimulation appeared intact. However, upregulation of one surface marker, B7.1, was impaired by the aly mutation (Fig. 1 C).

Bottom Line: Nuclear factor kappaB (NF-kappaB)-inducing kinase (NIK) has been implicated as a central transducing kinase in CD40-dependent activation.Here, we show that although NIK is essential for B cell activation, it is dispensable for activation of DCs.Such data provide compelling evidence that different intermediary kinases are used by different cellular lineages to trigger NF-kappaB activation via CD40.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Dartmouth Medical School, Lebanon, New Hampshire 03756, USA.

ABSTRACT
CD40 signaling in B cells and dendritic cells (DCs) is critical for the development of humoral and cell-mediated immunity, respectively. Nuclear factor kappaB (NF-kappaB)-inducing kinase (NIK) has been implicated as a central transducing kinase in CD40-dependent activation. Here, we show that although NIK is essential for B cell activation, it is dispensable for activation of DCs. Such data provide compelling evidence that different intermediary kinases are used by different cellular lineages to trigger NF-kappaB activation via CD40.

Show MeSH
Related in: MedlinePlus