MIMICKING THE TUMOR MICROENVIRONMENT OF CHRONIC LYMPHOCYTIC LEUKEMIA IN VITRO CRITICALLY DEPENDS ON THE TYPE OF B CELL RECEPTOR STIMULATION
(Abstract release date: 05/21/15)
EHA Library. Rombout A. 06/12/15; 102770; PB1710
Disclosure(s): Ghent University
Ans Rombout
Contributions
Contributions
Abstract
Abstract: PB1710
Type: Publication Only
Background
The B cell receptor (BCR) plays a key role in the crosstalk between chronic lymphocytic leukemia (CLL) cells and the tissue microenvironment, which favors disease progression by promoting proliferation and drug resistance. In these protective niches in the bone marrow and secondary lymphoid organs, the BCR becomes activated and sets a signaling cascade in motion that contributes to the survival and expansion of the malignant clone. In vitro studies of this crosstalk investigating downstream signaling and functional effects of BCR ligation often report contradictory results, in part due to the lack of a standardized protocol for BCR stimulation in CLL.
Aims
Our aim was to define a biologically relevant and robust in vitro stimulation method with regard to cellular phenotypic and transcriptional responses.
Methods
We evaluated mRNA (MYC, FOS, LPL) and protein (CD54, CD19, CD62L, CD184) expression of genes modulated by BCR triggering in IGHV mutated and unmutated CLL cells, after stimulation using soluble or immobilized anti-IgM antibodies from different brands.
Results
The effect of BCR stimulation on gene and protein expression was comparable in all CLL patients, irrespective of IGHV mutation status, as previously reported by us. However, immobilized anti-IgM stimulation elicited clear and robust changes in gene and protein expression, whereas the response to soluble anti-IgM was far less obvious.
Summary
These results show that the method of BCR stimulation is of major importance regarding responsiveness of CLL cells in the context of the tumor microenvironment, while genetic differences in the BCR pathway are less critical.
Session topic: Publication Only
Type: Publication Only
Background
The B cell receptor (BCR) plays a key role in the crosstalk between chronic lymphocytic leukemia (CLL) cells and the tissue microenvironment, which favors disease progression by promoting proliferation and drug resistance. In these protective niches in the bone marrow and secondary lymphoid organs, the BCR becomes activated and sets a signaling cascade in motion that contributes to the survival and expansion of the malignant clone. In vitro studies of this crosstalk investigating downstream signaling and functional effects of BCR ligation often report contradictory results, in part due to the lack of a standardized protocol for BCR stimulation in CLL.
Aims
Our aim was to define a biologically relevant and robust in vitro stimulation method with regard to cellular phenotypic and transcriptional responses.
Methods
We evaluated mRNA (MYC, FOS, LPL) and protein (CD54, CD19, CD62L, CD184) expression of genes modulated by BCR triggering in IGHV mutated and unmutated CLL cells, after stimulation using soluble or immobilized anti-IgM antibodies from different brands.
Results
The effect of BCR stimulation on gene and protein expression was comparable in all CLL patients, irrespective of IGHV mutation status, as previously reported by us. However, immobilized anti-IgM stimulation elicited clear and robust changes in gene and protein expression, whereas the response to soluble anti-IgM was far less obvious.
Summary
These results show that the method of BCR stimulation is of major importance regarding responsiveness of CLL cells in the context of the tumor microenvironment, while genetic differences in the BCR pathway are less critical.
Session topic: Publication Only
Abstract: PB1710
Type: Publication Only
Background
The B cell receptor (BCR) plays a key role in the crosstalk between chronic lymphocytic leukemia (CLL) cells and the tissue microenvironment, which favors disease progression by promoting proliferation and drug resistance. In these protective niches in the bone marrow and secondary lymphoid organs, the BCR becomes activated and sets a signaling cascade in motion that contributes to the survival and expansion of the malignant clone. In vitro studies of this crosstalk investigating downstream signaling and functional effects of BCR ligation often report contradictory results, in part due to the lack of a standardized protocol for BCR stimulation in CLL.
Aims
Our aim was to define a biologically relevant and robust in vitro stimulation method with regard to cellular phenotypic and transcriptional responses.
Methods
We evaluated mRNA (MYC, FOS, LPL) and protein (CD54, CD19, CD62L, CD184) expression of genes modulated by BCR triggering in IGHV mutated and unmutated CLL cells, after stimulation using soluble or immobilized anti-IgM antibodies from different brands.
Results
The effect of BCR stimulation on gene and protein expression was comparable in all CLL patients, irrespective of IGHV mutation status, as previously reported by us. However, immobilized anti-IgM stimulation elicited clear and robust changes in gene and protein expression, whereas the response to soluble anti-IgM was far less obvious.
Summary
These results show that the method of BCR stimulation is of major importance regarding responsiveness of CLL cells in the context of the tumor microenvironment, while genetic differences in the BCR pathway are less critical.
Session topic: Publication Only
Type: Publication Only
Background
The B cell receptor (BCR) plays a key role in the crosstalk between chronic lymphocytic leukemia (CLL) cells and the tissue microenvironment, which favors disease progression by promoting proliferation and drug resistance. In these protective niches in the bone marrow and secondary lymphoid organs, the BCR becomes activated and sets a signaling cascade in motion that contributes to the survival and expansion of the malignant clone. In vitro studies of this crosstalk investigating downstream signaling and functional effects of BCR ligation often report contradictory results, in part due to the lack of a standardized protocol for BCR stimulation in CLL.
Aims
Our aim was to define a biologically relevant and robust in vitro stimulation method with regard to cellular phenotypic and transcriptional responses.
Methods
We evaluated mRNA (MYC, FOS, LPL) and protein (CD54, CD19, CD62L, CD184) expression of genes modulated by BCR triggering in IGHV mutated and unmutated CLL cells, after stimulation using soluble or immobilized anti-IgM antibodies from different brands.
Results
The effect of BCR stimulation on gene and protein expression was comparable in all CLL patients, irrespective of IGHV mutation status, as previously reported by us. However, immobilized anti-IgM stimulation elicited clear and robust changes in gene and protein expression, whereas the response to soluble anti-IgM was far less obvious.
Summary
These results show that the method of BCR stimulation is of major importance regarding responsiveness of CLL cells in the context of the tumor microenvironment, while genetic differences in the BCR pathway are less critical.
Session topic: Publication Only
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