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Spinophilin participates in information transfer at immunological synapses.

Bloom O, Unternaehrer JJ, Jiang A, Shin JS, Delamarre L, Allen P, Mellman I - J. Cell Biol. (2008)

Bottom Line: In DCs interacting with T cells, spinophilin is polarized dynamically to contact sites in an antigen-dependent manner.It is also required for optimal T cell activation because DCs derived from mice lacking spinophilin exhibit defects in antigen presentation both in vitro and in vivo.Thus, spinophilin may play analogous roles in information transfer at both neuronal and immunological synapses.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology, Ludwig Institute for Cancer Research, New Haven, CT 06520, USA.

ABSTRACT
The adaptive immune response is initiated by the presentation of peptides bound to major histocompatibility complex molecules on dendritic cells (DCs) to antigen-specific T lymphocytes at a junction termed the immunological synapse. Although much attention has been paid to cytoplasmic events on the T cell side of the synapse, little is known concerning events on the DC side. We have sought signal transduction components of the neuronal synapse that were also expressed by DCs. One such protein is spinophilin, a scaffolding protein of neuronal dendritic spines that regulates synaptic transmission. In inactive, immature DCs, spinophilin is located throughout the cytoplasm but redistributes to the plasma membrane upon stimulus-induced maturation. In DCs interacting with T cells, spinophilin is polarized dynamically to contact sites in an antigen-dependent manner. It is also required for optimal T cell activation because DCs derived from mice lacking spinophilin exhibit defects in antigen presentation both in vitro and in vivo. Thus, spinophilin may play analogous roles in information transfer at both neuronal and immunological synapses.

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Spinophilin plays a functional role in antigen presentation in vivo. (a) Protocol: CD4+ T cells were isolated form TCR-transgenic mice (OT-II) and labeled with CFSE. 106 labeled cells were injected i.v. into WT and KO littermates. 1 d later, animals were injected i.v. with 10 μg ovalbumin + 100 ng LPS or LPS alone. On day three, spleen cells from WT and KO mice were isolated and analyzed for proliferation as measured by CFSE dilution or restimulated with ovalbumin (0, 10, and 20 μg/ml) for an additional 3 d. Intracellular cytokine staining was then performed. (b) The CD69hi population of adoptively transferred T cells was smaller in spinophilin KO (red) than in spinophilin WT (black) mice. Representative flow cytometry plots (left) and the data pooled from four independent experiments (right) is shown (n = 14 WT and 15 KO mice total; *, P < 0.05 by Student's t test). (c) The relative abundance of IFNγ-producing effector T cells was significantly greater in WT than in KO, as measured by flow cytometry of intracellular cytokine staining. (left) Representative flow cytometry plots. (right) Data pooled from three independent experiments (n = 8 animals total for WT and KO; ***, P < 0.001 by Student's t test).
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fig5: Spinophilin plays a functional role in antigen presentation in vivo. (a) Protocol: CD4+ T cells were isolated form TCR-transgenic mice (OT-II) and labeled with CFSE. 106 labeled cells were injected i.v. into WT and KO littermates. 1 d later, animals were injected i.v. with 10 μg ovalbumin + 100 ng LPS or LPS alone. On day three, spleen cells from WT and KO mice were isolated and analyzed for proliferation as measured by CFSE dilution or restimulated with ovalbumin (0, 10, and 20 μg/ml) for an additional 3 d. Intracellular cytokine staining was then performed. (b) The CD69hi population of adoptively transferred T cells was smaller in spinophilin KO (red) than in spinophilin WT (black) mice. Representative flow cytometry plots (left) and the data pooled from four independent experiments (right) is shown (n = 14 WT and 15 KO mice total; *, P < 0.05 by Student's t test). (c) The relative abundance of IFNγ-producing effector T cells was significantly greater in WT than in KO, as measured by flow cytometry of intracellular cytokine staining. (left) Representative flow cytometry plots. (right) Data pooled from three independent experiments (n = 8 animals total for WT and KO; ***, P < 0.001 by Student's t test).

Mentions: Given the diminished capacity of spinophilin KO DCs to present antigen in vitro, we next assayed the efficiency of antigen presentation in vivo by using naive antigen-specific T cells adoptively transferred into spinophilin KO and WT mice (Fig. 5 a). Mice were injected i.v. with 106 5(6)-carboxyfluorescein diacetate N-succinimidyl ester (CFSE)-labeled CD4+ T cells isolated from mice expressing an ovalbumin-specific TCR (OT-II). 24 h later, mice were injected i.v. with ovalbumin and LPS. After 2 d, spleens were isolated and the proliferation of adoptively transferred CD4+ T cells was measured by CFSE dilution. No significant defect in proliferation was observed (unpublished data). However, in the presence of an antigen, fewer CD4+ T cells adoptively transferred to spinophilin KO mice had elevated levels of CD69 than those transferred to WT littermates (Fig. 5 b), which suggests that spinophilin KO DCs are defective in activating T cells in vivo. As expected, there was no T cell activation in the absence of an antigen in either group of mice (unpublished data).


Spinophilin participates in information transfer at immunological synapses.

Bloom O, Unternaehrer JJ, Jiang A, Shin JS, Delamarre L, Allen P, Mellman I - J. Cell Biol. (2008)

Spinophilin plays a functional role in antigen presentation in vivo. (a) Protocol: CD4+ T cells were isolated form TCR-transgenic mice (OT-II) and labeled with CFSE. 106 labeled cells were injected i.v. into WT and KO littermates. 1 d later, animals were injected i.v. with 10 μg ovalbumin + 100 ng LPS or LPS alone. On day three, spleen cells from WT and KO mice were isolated and analyzed for proliferation as measured by CFSE dilution or restimulated with ovalbumin (0, 10, and 20 μg/ml) for an additional 3 d. Intracellular cytokine staining was then performed. (b) The CD69hi population of adoptively transferred T cells was smaller in spinophilin KO (red) than in spinophilin WT (black) mice. Representative flow cytometry plots (left) and the data pooled from four independent experiments (right) is shown (n = 14 WT and 15 KO mice total; *, P < 0.05 by Student's t test). (c) The relative abundance of IFNγ-producing effector T cells was significantly greater in WT than in KO, as measured by flow cytometry of intracellular cytokine staining. (left) Representative flow cytometry plots. (right) Data pooled from three independent experiments (n = 8 animals total for WT and KO; ***, P < 0.001 by Student's t test).
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fig5: Spinophilin plays a functional role in antigen presentation in vivo. (a) Protocol: CD4+ T cells were isolated form TCR-transgenic mice (OT-II) and labeled with CFSE. 106 labeled cells were injected i.v. into WT and KO littermates. 1 d later, animals were injected i.v. with 10 μg ovalbumin + 100 ng LPS or LPS alone. On day three, spleen cells from WT and KO mice were isolated and analyzed for proliferation as measured by CFSE dilution or restimulated with ovalbumin (0, 10, and 20 μg/ml) for an additional 3 d. Intracellular cytokine staining was then performed. (b) The CD69hi population of adoptively transferred T cells was smaller in spinophilin KO (red) than in spinophilin WT (black) mice. Representative flow cytometry plots (left) and the data pooled from four independent experiments (right) is shown (n = 14 WT and 15 KO mice total; *, P < 0.05 by Student's t test). (c) The relative abundance of IFNγ-producing effector T cells was significantly greater in WT than in KO, as measured by flow cytometry of intracellular cytokine staining. (left) Representative flow cytometry plots. (right) Data pooled from three independent experiments (n = 8 animals total for WT and KO; ***, P < 0.001 by Student's t test).
Mentions: Given the diminished capacity of spinophilin KO DCs to present antigen in vitro, we next assayed the efficiency of antigen presentation in vivo by using naive antigen-specific T cells adoptively transferred into spinophilin KO and WT mice (Fig. 5 a). Mice were injected i.v. with 106 5(6)-carboxyfluorescein diacetate N-succinimidyl ester (CFSE)-labeled CD4+ T cells isolated from mice expressing an ovalbumin-specific TCR (OT-II). 24 h later, mice were injected i.v. with ovalbumin and LPS. After 2 d, spleens were isolated and the proliferation of adoptively transferred CD4+ T cells was measured by CFSE dilution. No significant defect in proliferation was observed (unpublished data). However, in the presence of an antigen, fewer CD4+ T cells adoptively transferred to spinophilin KO mice had elevated levels of CD69 than those transferred to WT littermates (Fig. 5 b), which suggests that spinophilin KO DCs are defective in activating T cells in vivo. As expected, there was no T cell activation in the absence of an antigen in either group of mice (unpublished data).

Bottom Line: In DCs interacting with T cells, spinophilin is polarized dynamically to contact sites in an antigen-dependent manner.It is also required for optimal T cell activation because DCs derived from mice lacking spinophilin exhibit defects in antigen presentation both in vitro and in vivo.Thus, spinophilin may play analogous roles in information transfer at both neuronal and immunological synapses.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology, Ludwig Institute for Cancer Research, New Haven, CT 06520, USA.

ABSTRACT
The adaptive immune response is initiated by the presentation of peptides bound to major histocompatibility complex molecules on dendritic cells (DCs) to antigen-specific T lymphocytes at a junction termed the immunological synapse. Although much attention has been paid to cytoplasmic events on the T cell side of the synapse, little is known concerning events on the DC side. We have sought signal transduction components of the neuronal synapse that were also expressed by DCs. One such protein is spinophilin, a scaffolding protein of neuronal dendritic spines that regulates synaptic transmission. In inactive, immature DCs, spinophilin is located throughout the cytoplasm but redistributes to the plasma membrane upon stimulus-induced maturation. In DCs interacting with T cells, spinophilin is polarized dynamically to contact sites in an antigen-dependent manner. It is also required for optimal T cell activation because DCs derived from mice lacking spinophilin exhibit defects in antigen presentation both in vitro and in vivo. Thus, spinophilin may play analogous roles in information transfer at both neuronal and immunological synapses.

Show MeSH
Related in: MedlinePlus