IL4 and CD40L Prevent Apoptosis of Chronic Lymphocytic Leukemia Cells Via Intracellular pSTAT6 and NFkB Signaling and Not Via Receptor Kinetics

Gespeichert in:

Bibliographische Detailangaben
Zeitschriftentitel:	Blood
Personen und Körperschaften:	Mertens, Daniel, Bhattacharya, Nupur, Häbe, Sarah, Döhner, Hartmut, Stilgenbauer, Stephan
In:	Blood, 120, 2012, 21, S. 3893-3893
Format:	E-Article
Sprache:	Englisch
veröffentlicht:	American Society of Hematology
Schlagwörter:	Cell Biology Hematology Immunology Biochemistry

author_facet	Mertens, Daniel Bhattacharya, Nupur Häbe, Sarah Döhner, Hartmut Stilgenbauer, Stephan Mertens, Daniel Bhattacharya, Nupur Häbe, Sarah Döhner, Hartmut Stilgenbauer, Stephan
author	Mertens, Daniel Bhattacharya, Nupur Häbe, Sarah Döhner, Hartmut Stilgenbauer, Stephan
spellingShingle	Mertens, Daniel Bhattacharya, Nupur Häbe, Sarah Döhner, Hartmut Stilgenbauer, Stephan Blood IL4 and CD40L Prevent Apoptosis of Chronic Lymphocytic Leukemia Cells Via Intracellular pSTAT6 and NFkB Signaling and Not Via Receptor Kinetics Cell Biology Hematology Immunology Biochemistry
author_sort	mertens, daniel
spelling	Mertens, Daniel Bhattacharya, Nupur Häbe, Sarah Döhner, Hartmut Stilgenbauer, Stephan 0006-4971 1528-0020 American Society of Hematology Cell Biology Hematology Immunology Biochemistry http://dx.doi.org/10.1182/blood.v120.21.3893.3893 <jats:title>Abstract</jats:title> <jats:p>Abstract 3893</jats:p> <jats:p>Chronic lymphocytic leukemia (CLL) cells are highly dependent on microenvironmental input for their extended survival in vivo, but the underlying molecular mechanism is still unclear. Compared to non-malignant B-cells, CLL cells are more responsive to contact dependent complex stimuli like coculture on bone marrow derived stromal cell lines of both human (p<0.0001) and murine origin (p<0.01), but also to soluble factors (human conditioned medium p<0.0001, murine conditioned medium p<0.001, all student′s t-test). In order to understand the intrinsic difference of the anti-apoptotic phenotype of CLL cells, the signalling circuitry of the malignant cells was modelled. Compared to candidate ligands like SDF-1 (at concentrations between 10–1000ng/ml), BAFF (250–1000ng/ml), APRIL (250–1000ng/ml) and soluble anti-IgM (1–25μg/ml), the factors CD40L (10–2000ng/ml) and IL4 (0.1–10ng/ml) were the most efficient ligands in rescuing CLL cells from spontaneous death in vitro. The dose response of IL4 and CD40L displayed different saturation and cooperativity between CLL cells and non-malignant B-cells. Using IL4, saturation was reached both for CLL cells and B-cells at 0.2pM, but at 52% survival (+/− 8%) for CLL cells and 28% (+/−7%) for B-cells, and the estimated dissociation constant Kd was 0.01pM for both ligands. For CD40L, CLL cell survival reached saturation at 40nM, while no saturation was reached for B-cells. Intriguingly, B-cells showed cooperativity in their response to CD40L, with a cooperativity coefficient of 2.0 and a Kd of 70pM, while cooperativity for CD40L was lost in CLL cells (Kd of only 2.6pM). This pointed towards distinct differences in ligand-receptor interactions or in downstream signaling between CLL cells and non-malignant B-cells. However, high-throughput spatial analysis with a microscope-coupled cytometer did not show differences of receptor quantity or receptor distribution between malignant and non-malignant cells. In contrast, quantity and phosphorylation levels of downstream signalling nodes like STAT6 (measured by flow cytometry and validated by Western-blot) and the activity of NF-kB (p65 binding to DNA measured by oligonucleotide-coupled ELISA) were higher in CLL cells compared to B-cells from healthy donors. Therefore, the defect in IL4 and CD40L signalling that leads to an enhanced survival in CLL cells is likely caused by changes in the intracellular circuitry.</jats:p> <jats:sec> <jats:title>Disclosures:</jats:title> <jats:p>No relevant conflicts of interest to declare.</jats:p> </jats:sec> IL4 and CD40L Prevent Apoptosis of Chronic Lymphocytic Leukemia Cells Via Intracellular pSTAT6 and NFkB Signaling and Not Via Receptor Kinetics Blood
doi_str_mv	10.1182/blood.v120.21.3893.3893
facet_avail	Online Free
finc_class_facet	Biologie Medizin Chemie und Pharmazie
format	ElectronicArticle
fullrecord	blob:ai-49-aHR0cDovL2R4LmRvaS5vcmcvMTAuMTE4Mi9ibG9vZC52MTIwLjIxLjM4OTMuMzg5Mw
id	ai-49-aHR0cDovL2R4LmRvaS5vcmcvMTAuMTE4Mi9ibG9vZC52MTIwLjIxLjM4OTMuMzg5Mw
institution	DE-Ch1 DE-L229 DE-D275 DE-Bn3 DE-Brt1 DE-Zwi2 DE-D161 DE-Gla1 DE-Zi4 DE-15 DE-Pl11 DE-Rs1 DE-105 DE-14
imprint	American Society of Hematology, 2012
imprint_str_mv	American Society of Hematology, 2012
issn	0006-4971 1528-0020
issn_str_mv	0006-4971 1528-0020
language	English
mega_collection	American Society of Hematology (CrossRef)
match_str	mertens2012il4andcd40lpreventapoptosisofchroniclymphocyticleukemiacellsviaintracellularpstat6andnfkbsignalingandnotviareceptorkinetics
publishDateSort	2012
publisher	American Society of Hematology
recordtype	ai
record_format	ai
series	Blood
source_id	49
title	IL4 and CD40L Prevent Apoptosis of Chronic Lymphocytic Leukemia Cells Via Intracellular pSTAT6 and NFkB Signaling and Not Via Receptor Kinetics
title_unstemmed	IL4 and CD40L Prevent Apoptosis of Chronic Lymphocytic Leukemia Cells Via Intracellular pSTAT6 and NFkB Signaling and Not Via Receptor Kinetics
title_full	IL4 and CD40L Prevent Apoptosis of Chronic Lymphocytic Leukemia Cells Via Intracellular pSTAT6 and NFkB Signaling and Not Via Receptor Kinetics
title_fullStr	IL4 and CD40L Prevent Apoptosis of Chronic Lymphocytic Leukemia Cells Via Intracellular pSTAT6 and NFkB Signaling and Not Via Receptor Kinetics
title_full_unstemmed	IL4 and CD40L Prevent Apoptosis of Chronic Lymphocytic Leukemia Cells Via Intracellular pSTAT6 and NFkB Signaling and Not Via Receptor Kinetics
title_short	IL4 and CD40L Prevent Apoptosis of Chronic Lymphocytic Leukemia Cells Via Intracellular pSTAT6 and NFkB Signaling and Not Via Receptor Kinetics
title_sort	il4 and cd40l prevent apoptosis of chronic lymphocytic leukemia cells via intracellular pstat6 and nfkb signaling and not via receptor kinetics
topic	Cell Biology Hematology Immunology Biochemistry
url	http://dx.doi.org/10.1182/blood.v120.21.3893.3893
publishDate	2012
physical	3893-3893
description	<jats:title>Abstract</jats:title> <jats:p>Abstract 3893</jats:p> <jats:p>Chronic lymphocytic leukemia (CLL) cells are highly dependent on microenvironmental input for their extended survival in vivo, but the underlying molecular mechanism is still unclear. Compared to non-malignant B-cells, CLL cells are more responsive to contact dependent complex stimuli like coculture on bone marrow derived stromal cell lines of both human (p<0.0001) and murine origin (p<0.01), but also to soluble factors (human conditioned medium p<0.0001, murine conditioned medium p<0.001, all student′s t-test). In order to understand the intrinsic difference of the anti-apoptotic phenotype of CLL cells, the signalling circuitry of the malignant cells was modelled. Compared to candidate ligands like SDF-1 (at concentrations between 10–1000ng/ml), BAFF (250–1000ng/ml), APRIL (250–1000ng/ml) and soluble anti-IgM (1–25μg/ml), the factors CD40L (10–2000ng/ml) and IL4 (0.1–10ng/ml) were the most efficient ligands in rescuing CLL cells from spontaneous death in vitro. The dose response of IL4 and CD40L displayed different saturation and cooperativity between CLL cells and non-malignant B-cells. Using IL4, saturation was reached both for CLL cells and B-cells at 0.2pM, but at 52% survival (+/− 8%) for CLL cells and 28% (+/−7%) for B-cells, and the estimated dissociation constant Kd was 0.01pM for both ligands. For CD40L, CLL cell survival reached saturation at 40nM, while no saturation was reached for B-cells. Intriguingly, B-cells showed cooperativity in their response to CD40L, with a cooperativity coefficient of 2.0 and a Kd of 70pM, while cooperativity for CD40L was lost in CLL cells (Kd of only 2.6pM). This pointed towards distinct differences in ligand-receptor interactions or in downstream signaling between CLL cells and non-malignant B-cells. However, high-throughput spatial analysis with a microscope-coupled cytometer did not show differences of receptor quantity or receptor distribution between malignant and non-malignant cells. In contrast, quantity and phosphorylation levels of downstream signalling nodes like STAT6 (measured by flow cytometry and validated by Western-blot) and the activity of NF-kB (p65 binding to DNA measured by oligonucleotide-coupled ELISA) were higher in CLL cells compared to B-cells from healthy donors. Therefore, the defect in IL4 and CD40L signalling that leads to an enhanced survival in CLL cells is likely caused by changes in the intracellular circuitry.</jats:p> <jats:sec> <jats:title>Disclosures:</jats:title> <jats:p>No relevant conflicts of interest to declare.</jats:p> </jats:sec>
container_issue	21
container_start_page	3893
container_title	Blood
container_volume	120
format_de105	Article, E-Article
format_de14	Article, E-Article
format_de15	Article, E-Article
format_de520	Article, E-Article
format_de540	Article, E-Article
format_dech1	Article, E-Article
format_ded117	Article, E-Article
format_degla1	E-Article
format_del152	Buch
format_del189	Article, E-Article
format_dezi4	Article
format_dezwi2	Article, E-Article
format_finc	Article, E-Article
format_nrw	Article, E-Article
_version_	1792325251284074499
geogr_code	not assigned
last_indexed	2024-03-01T12:02:37.942Z
geogr_code_person	not assigned
openURL	url_ver=Z39.88-2004&ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fvufind.svn.sourceforge.net%3Agenerator&rft.title=IL4+and+CD40L+Prevent+Apoptosis+of+Chronic+Lymphocytic+Leukemia+Cells+Via+Intracellular+pSTAT6+and+NFkB+Signaling+and+Not+Via+Receptor+Kinetics&rft.date=2012-11-16&genre=article&issn=1528-0020&volume=120&issue=21&spage=3893&epage=3893&pages=3893-3893&jtitle=Blood&atitle=IL4+and+CD40L+Prevent+Apoptosis+of+Chronic+Lymphocytic+Leukemia+Cells+Via+Intracellular+pSTAT6+and+NFkB+Signaling+and+Not+Via+Receptor+Kinetics&aulast=Stilgenbauer&aufirst=Stephan&rft_id=info%3Adoi%2F10.1182%2Fblood.v120.21.3893.3893&rft.language%5B0%5D=eng
SOLR
_version_	1792325251284074499
author	Mertens, Daniel, Bhattacharya, Nupur, Häbe, Sarah, Döhner, Hartmut, Stilgenbauer, Stephan
author_facet	Mertens, Daniel, Bhattacharya, Nupur, Häbe, Sarah, Döhner, Hartmut, Stilgenbauer, Stephan, Mertens, Daniel, Bhattacharya, Nupur, Häbe, Sarah, Döhner, Hartmut, Stilgenbauer, Stephan
author_sort	mertens, daniel
container_issue	21
container_start_page	3893
container_title	Blood
container_volume	120
description	<jats:title>Abstract</jats:title> <jats:p>Abstract 3893</jats:p> <jats:p>Chronic lymphocytic leukemia (CLL) cells are highly dependent on microenvironmental input for their extended survival in vivo, but the underlying molecular mechanism is still unclear. Compared to non-malignant B-cells, CLL cells are more responsive to contact dependent complex stimuli like coculture on bone marrow derived stromal cell lines of both human (p<0.0001) and murine origin (p<0.01), but also to soluble factors (human conditioned medium p<0.0001, murine conditioned medium p<0.001, all student′s t-test). In order to understand the intrinsic difference of the anti-apoptotic phenotype of CLL cells, the signalling circuitry of the malignant cells was modelled. Compared to candidate ligands like SDF-1 (at concentrations between 10–1000ng/ml), BAFF (250–1000ng/ml), APRIL (250–1000ng/ml) and soluble anti-IgM (1–25μg/ml), the factors CD40L (10–2000ng/ml) and IL4 (0.1–10ng/ml) were the most efficient ligands in rescuing CLL cells from spontaneous death in vitro. The dose response of IL4 and CD40L displayed different saturation and cooperativity between CLL cells and non-malignant B-cells. Using IL4, saturation was reached both for CLL cells and B-cells at 0.2pM, but at 52% survival (+/− 8%) for CLL cells and 28% (+/−7%) for B-cells, and the estimated dissociation constant Kd was 0.01pM for both ligands. For CD40L, CLL cell survival reached saturation at 40nM, while no saturation was reached for B-cells. Intriguingly, B-cells showed cooperativity in their response to CD40L, with a cooperativity coefficient of 2.0 and a Kd of 70pM, while cooperativity for CD40L was lost in CLL cells (Kd of only 2.6pM). This pointed towards distinct differences in ligand-receptor interactions or in downstream signaling between CLL cells and non-malignant B-cells. However, high-throughput spatial analysis with a microscope-coupled cytometer did not show differences of receptor quantity or receptor distribution between malignant and non-malignant cells. In contrast, quantity and phosphorylation levels of downstream signalling nodes like STAT6 (measured by flow cytometry and validated by Western-blot) and the activity of NF-kB (p65 binding to DNA measured by oligonucleotide-coupled ELISA) were higher in CLL cells compared to B-cells from healthy donors. Therefore, the defect in IL4 and CD40L signalling that leads to an enhanced survival in CLL cells is likely caused by changes in the intracellular circuitry.</jats:p> <jats:sec> <jats:title>Disclosures:</jats:title> <jats:p>No relevant conflicts of interest to declare.</jats:p> </jats:sec>
doi_str_mv	10.1182/blood.v120.21.3893.3893
facet_avail	Online, Free
finc_class_facet	Biologie, Medizin, Chemie und Pharmazie
format	ElectronicArticle
format_de105	Article, E-Article
format_de14	Article, E-Article
format_de15	Article, E-Article
format_de520	Article, E-Article
format_de540	Article, E-Article
format_dech1	Article, E-Article
format_ded117	Article, E-Article
format_degla1	E-Article
format_del152	Buch
format_del189	Article, E-Article
format_dezi4	Article
format_dezwi2	Article, E-Article
format_finc	Article, E-Article
format_nrw	Article, E-Article
geogr_code	not assigned
geogr_code_person	not assigned
id	ai-49-aHR0cDovL2R4LmRvaS5vcmcvMTAuMTE4Mi9ibG9vZC52MTIwLjIxLjM4OTMuMzg5Mw
imprint	American Society of Hematology, 2012
imprint_str_mv	American Society of Hematology, 2012
institution	DE-Ch1, DE-L229, DE-D275, DE-Bn3, DE-Brt1, DE-Zwi2, DE-D161, DE-Gla1, DE-Zi4, DE-15, DE-Pl11, DE-Rs1, DE-105, DE-14
issn	0006-4971, 1528-0020
issn_str_mv	0006-4971, 1528-0020
language	English
last_indexed	2024-03-01T12:02:37.942Z
match_str	mertens2012il4andcd40lpreventapoptosisofchroniclymphocyticleukemiacellsviaintracellularpstat6andnfkbsignalingandnotviareceptorkinetics
mega_collection	American Society of Hematology (CrossRef)
physical	3893-3893
publishDate	2012
publishDateSort	2012
publisher	American Society of Hematology
record_format	ai
recordtype	ai
series	Blood
source_id	49
spelling	Mertens, Daniel Bhattacharya, Nupur Häbe, Sarah Döhner, Hartmut Stilgenbauer, Stephan 0006-4971 1528-0020 American Society of Hematology Cell Biology Hematology Immunology Biochemistry http://dx.doi.org/10.1182/blood.v120.21.3893.3893 <jats:title>Abstract</jats:title> <jats:p>Abstract 3893</jats:p> <jats:p>Chronic lymphocytic leukemia (CLL) cells are highly dependent on microenvironmental input for their extended survival in vivo, but the underlying molecular mechanism is still unclear. Compared to non-malignant B-cells, CLL cells are more responsive to contact dependent complex stimuli like coculture on bone marrow derived stromal cell lines of both human (p<0.0001) and murine origin (p<0.01), but also to soluble factors (human conditioned medium p<0.0001, murine conditioned medium p<0.001, all student′s t-test). In order to understand the intrinsic difference of the anti-apoptotic phenotype of CLL cells, the signalling circuitry of the malignant cells was modelled. Compared to candidate ligands like SDF-1 (at concentrations between 10–1000ng/ml), BAFF (250–1000ng/ml), APRIL (250–1000ng/ml) and soluble anti-IgM (1–25μg/ml), the factors CD40L (10–2000ng/ml) and IL4 (0.1–10ng/ml) were the most efficient ligands in rescuing CLL cells from spontaneous death in vitro. The dose response of IL4 and CD40L displayed different saturation and cooperativity between CLL cells and non-malignant B-cells. Using IL4, saturation was reached both for CLL cells and B-cells at 0.2pM, but at 52% survival (+/− 8%) for CLL cells and 28% (+/−7%) for B-cells, and the estimated dissociation constant Kd was 0.01pM for both ligands. For CD40L, CLL cell survival reached saturation at 40nM, while no saturation was reached for B-cells. Intriguingly, B-cells showed cooperativity in their response to CD40L, with a cooperativity coefficient of 2.0 and a Kd of 70pM, while cooperativity for CD40L was lost in CLL cells (Kd of only 2.6pM). This pointed towards distinct differences in ligand-receptor interactions or in downstream signaling between CLL cells and non-malignant B-cells. However, high-throughput spatial analysis with a microscope-coupled cytometer did not show differences of receptor quantity or receptor distribution between malignant and non-malignant cells. In contrast, quantity and phosphorylation levels of downstream signalling nodes like STAT6 (measured by flow cytometry and validated by Western-blot) and the activity of NF-kB (p65 binding to DNA measured by oligonucleotide-coupled ELISA) were higher in CLL cells compared to B-cells from healthy donors. Therefore, the defect in IL4 and CD40L signalling that leads to an enhanced survival in CLL cells is likely caused by changes in the intracellular circuitry.</jats:p> <jats:sec> <jats:title>Disclosures:</jats:title> <jats:p>No relevant conflicts of interest to declare.</jats:p> </jats:sec> IL4 and CD40L Prevent Apoptosis of Chronic Lymphocytic Leukemia Cells Via Intracellular pSTAT6 and NFkB Signaling and Not Via Receptor Kinetics Blood
spellingShingle	Mertens, Daniel, Bhattacharya, Nupur, Häbe, Sarah, Döhner, Hartmut, Stilgenbauer, Stephan, Blood, IL4 and CD40L Prevent Apoptosis of Chronic Lymphocytic Leukemia Cells Via Intracellular pSTAT6 and NFkB Signaling and Not Via Receptor Kinetics, Cell Biology, Hematology, Immunology, Biochemistry
title	IL4 and CD40L Prevent Apoptosis of Chronic Lymphocytic Leukemia Cells Via Intracellular pSTAT6 and NFkB Signaling and Not Via Receptor Kinetics
title_full	IL4 and CD40L Prevent Apoptosis of Chronic Lymphocytic Leukemia Cells Via Intracellular pSTAT6 and NFkB Signaling and Not Via Receptor Kinetics
title_fullStr	IL4 and CD40L Prevent Apoptosis of Chronic Lymphocytic Leukemia Cells Via Intracellular pSTAT6 and NFkB Signaling and Not Via Receptor Kinetics
title_full_unstemmed	IL4 and CD40L Prevent Apoptosis of Chronic Lymphocytic Leukemia Cells Via Intracellular pSTAT6 and NFkB Signaling and Not Via Receptor Kinetics
title_short	IL4 and CD40L Prevent Apoptosis of Chronic Lymphocytic Leukemia Cells Via Intracellular pSTAT6 and NFkB Signaling and Not Via Receptor Kinetics
title_sort	il4 and cd40l prevent apoptosis of chronic lymphocytic leukemia cells via intracellular pstat6 and nfkb signaling and not via receptor kinetics
title_unstemmed	IL4 and CD40L Prevent Apoptosis of Chronic Lymphocytic Leukemia Cells Via Intracellular pSTAT6 and NFkB Signaling and Not Via Receptor Kinetics
topic	Cell Biology, Hematology, Immunology, Biochemistry
url	http://dx.doi.org/10.1182/blood.v120.21.3893.3893