Regulation of NKG2D-ligand cell surface expression by intracellular calcium after HDAC-inhibitor treatment

Helle Jensen; Michael Henrik Hagemann-Jensen; Felicia Kathrine Bratt Lauridsen; Søren Skov

doi:10.1016/j.molimm.2012.08.011

Regulation of NKG2D-ligand cell surface expression by intracellular calcium after HDAC-inhibitor treatment

Helle Jensen, Michael Henrik Hagemann-Jensen, Felicia Kathrine Bratt Lauridsen, Søren Skov

Department of Biomedical Sciences

7 Citations (Scopus)

Abstract

In this study we demonstrate that histone deacetylase (HDAC)-inhibitor mediated cell surface expression of the structural different NKG2D-ligands MICA/B and ULBP2 is calcium-dependent. Treatment with the calcium chelator EGTA inhibited constitutive as well as HDAC-inhibitor induced MICA/B and ULBP2 cell surface expression on melanoma cells and Jurkat T-cells. A NKG2D-dependent cytolytic assay and staining with a recombinant NKG2D-Fc fusion protein showed that calcium chelation impaired the functional ability of NKG2D-ligands induced by HDAC-inhibitor treatment. The HDAC-inhibitor induced cell surface expression of ULBP2, but not MICA/B, was sensitive to treatment calmidazolium and trifluoperazine, two agents known to block calcium signaling. siRNA-mediated knock-down of the calcium-regulated proteins calmodulin or calpain did however not affect NKG2D-ligand cell surface expression on Jurkat T-cells. We further show that secretion and cell surface binding of the calcium-regulating protein galectin-1 is enhanced upon HDAC-inhibitor treatment of melanoma cells. However, binding of galectin-1 to cell surface glycoproteins was not critical for constitutive or HDAC-inhibitor induced MICA/B and ULBP2 cell surface expression. We provide evidence that MICA/B and ULBP2 cell surface expression is controlled differently by calcium, which adds to the increasing perception that cell surface expression of MICA/B and ULBP2 is controlled by distinct signal transduction pathways.

Original language	English
Journal	Molecular Immunology
Volume	53
Issue number	3
Pages (from-to)	255-264
Number of pages	10
ISSN	0161-5890
DOIs	https://doi.org/10.1016/j.molimm.2012.08.011
Publication status	Published - Mar 2013

Keywords

Base Sequence
Calcium Signaling
Calmodulin
Calpain
Cell Line, Tumor
Cell Membrane
Depsipeptides
GPI-Linked Proteins
Galectin 1
Gene Knockdown Techniques
Histone Deacetylase Inhibitors
Humans
Imidazoles
Intercellular Signaling Peptides and Proteins
Jurkat Cells
Ligands
Melanoma
RNA, Small Interfering
Trifluoperazine

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10.1016/j.molimm.2012.08.011

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title = "Regulation of NKG2D-ligand cell surface expression by intracellular calcium after HDAC-inhibitor treatment",

abstract = "In this study we demonstrate that histone deacetylase (HDAC)-inhibitor mediated cell surface expression of the structural different NKG2D-ligands MICA/B and ULBP2 is calcium-dependent. Treatment with the calcium chelator EGTA inhibited constitutive as well as HDAC-inhibitor induced MICA/B and ULBP2 cell surface expression on melanoma cells and Jurkat T-cells. A NKG2D-dependent cytolytic assay and staining with a recombinant NKG2D-Fc fusion protein showed that calcium chelation impaired the functional ability of NKG2D-ligands induced by HDAC-inhibitor treatment. The HDAC-inhibitor induced cell surface expression of ULBP2, but not MICA/B, was sensitive to treatment calmidazolium and trifluoperazine, two agents known to block calcium signaling. siRNA-mediated knock-down of the calcium-regulated proteins calmodulin or calpain did however not affect NKG2D-ligand cell surface expression on Jurkat T-cells. We further show that secretion and cell surface binding of the calcium-regulating protein galectin-1 is enhanced upon HDAC-inhibitor treatment of melanoma cells. However, binding of galectin-1 to cell surface glycoproteins was not critical for constitutive or HDAC-inhibitor induced MICA/B and ULBP2 cell surface expression. We provide evidence that MICA/B and ULBP2 cell surface expression is controlled differently by calcium, which adds to the increasing perception that cell surface expression of MICA/B and ULBP2 is controlled by distinct signal transduction pathways.",

keywords = "Base Sequence, Calcium Signaling, Calmodulin, Calpain, Cell Line, Tumor, Cell Membrane, Depsipeptides, GPI-Linked Proteins, Galectin 1, Gene Knockdown Techniques, Histone Deacetylase Inhibitors, Humans, Imidazoles, Intercellular Signaling Peptides and Proteins, Jurkat Cells, Ligands, Melanoma, RNA, Small Interfering, Trifluoperazine, NKG2D-ligand, ULBP2, Calcium, Galectin-1, HDAC-inhibitor, Cancer",

author = "Helle Jensen and Hagemann-Jensen, {Michael Henrik} and Lauridsen, {Felicia Kathrine Bratt} and S{\o}ren Skov",

year = "2013",

month = mar,

doi = "10.1016/j.molimm.2012.08.011",

language = "English",

volume = "53",

pages = "255--264",

journal = "Molecular Immunology",

issn = "0161-5890",

publisher = "Pergamon Press",

number = "3",

}

TY - JOUR

T1 - Regulation of NKG2D-ligand cell surface expression by intracellular calcium after HDAC-inhibitor treatment

AU - Jensen, Helle

AU - Hagemann-Jensen, Michael Henrik

AU - Lauridsen, Felicia Kathrine Bratt

AU - Skov, Søren

PY - 2013/3

Y1 - 2013/3

N2 - In this study we demonstrate that histone deacetylase (HDAC)-inhibitor mediated cell surface expression of the structural different NKG2D-ligands MICA/B and ULBP2 is calcium-dependent. Treatment with the calcium chelator EGTA inhibited constitutive as well as HDAC-inhibitor induced MICA/B and ULBP2 cell surface expression on melanoma cells and Jurkat T-cells. A NKG2D-dependent cytolytic assay and staining with a recombinant NKG2D-Fc fusion protein showed that calcium chelation impaired the functional ability of NKG2D-ligands induced by HDAC-inhibitor treatment. The HDAC-inhibitor induced cell surface expression of ULBP2, but not MICA/B, was sensitive to treatment calmidazolium and trifluoperazine, two agents known to block calcium signaling. siRNA-mediated knock-down of the calcium-regulated proteins calmodulin or calpain did however not affect NKG2D-ligand cell surface expression on Jurkat T-cells. We further show that secretion and cell surface binding of the calcium-regulating protein galectin-1 is enhanced upon HDAC-inhibitor treatment of melanoma cells. However, binding of galectin-1 to cell surface glycoproteins was not critical for constitutive or HDAC-inhibitor induced MICA/B and ULBP2 cell surface expression. We provide evidence that MICA/B and ULBP2 cell surface expression is controlled differently by calcium, which adds to the increasing perception that cell surface expression of MICA/B and ULBP2 is controlled by distinct signal transduction pathways.

AB - In this study we demonstrate that histone deacetylase (HDAC)-inhibitor mediated cell surface expression of the structural different NKG2D-ligands MICA/B and ULBP2 is calcium-dependent. Treatment with the calcium chelator EGTA inhibited constitutive as well as HDAC-inhibitor induced MICA/B and ULBP2 cell surface expression on melanoma cells and Jurkat T-cells. A NKG2D-dependent cytolytic assay and staining with a recombinant NKG2D-Fc fusion protein showed that calcium chelation impaired the functional ability of NKG2D-ligands induced by HDAC-inhibitor treatment. The HDAC-inhibitor induced cell surface expression of ULBP2, but not MICA/B, was sensitive to treatment calmidazolium and trifluoperazine, two agents known to block calcium signaling. siRNA-mediated knock-down of the calcium-regulated proteins calmodulin or calpain did however not affect NKG2D-ligand cell surface expression on Jurkat T-cells. We further show that secretion and cell surface binding of the calcium-regulating protein galectin-1 is enhanced upon HDAC-inhibitor treatment of melanoma cells. However, binding of galectin-1 to cell surface glycoproteins was not critical for constitutive or HDAC-inhibitor induced MICA/B and ULBP2 cell surface expression. We provide evidence that MICA/B and ULBP2 cell surface expression is controlled differently by calcium, which adds to the increasing perception that cell surface expression of MICA/B and ULBP2 is controlled by distinct signal transduction pathways.

KW - Base Sequence

KW - Calcium Signaling

KW - Calmodulin

KW - Calpain

KW - Cell Line, Tumor

KW - Cell Membrane

KW - Depsipeptides

KW - GPI-Linked Proteins

KW - Galectin 1

KW - Gene Knockdown Techniques

KW - Histone Deacetylase Inhibitors

KW - Humans

KW - Imidazoles

KW - Intercellular Signaling Peptides and Proteins

KW - Jurkat Cells

KW - Ligands

KW - Melanoma

KW - RNA, Small Interfering

KW - Trifluoperazine

KW - NKG2D-ligand

KW - ULBP2

KW - Calcium

KW - Galectin-1

KW - HDAC-inhibitor

KW - Cancer

U2 - 10.1016/j.molimm.2012.08.011

DO - 10.1016/j.molimm.2012.08.011

M3 - Journal article

C2 - 22964480

SN - 0161-5890

VL - 53

SP - 255

EP - 264

JO - Molecular Immunology

JF - Molecular Immunology

IS - 3

ER -

Regulation of NKG2D-ligand cell surface expression by intracellular calcium after HDAC-inhibitor treatment

Abstract

Keywords

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