Synergy of interleukin 10 and toll-like receptor 9 signalling in B cell proliferation: Implications for lymphoma pathogenesis.
A network of autocrine and paracrine signals defines B cell homeostasis and is thought to be involved in transformation processes. Investigating interactions of these microenvironmental factors and their relation to proto-oncogenes as c-Myc (MYC) is fundamental to understand the biology of B cell lymphoma. Therefore, B cells with conditional MYC expression were stimulated with CD40L, insulin-like growth factor 1, α-IgM, Interleukin-10 (IL10) and CpG alone or in combination. The impact of forty different interventions on cell proliferation was investigated in MYC deprived cells and calculated by linear regression. Combination of CpG and IL10 led to a strong synergistic activation of cell proliferation (S-phase/doubling of total cell number) comparable to cells with high MYC expression. A synergistic up-regulation of CDK4, CDK6 and CCND3 expression by IL10 and CpG treatment was causal for this proliferative effect as shown by qRT-PCR analysis and inhibition of the CDK4/6 complex by PD0332991. Furthermore, treatment of stimulated MYC deprived cells with MLN120b, ACHP, Pyridone 6 or Ruxolitinib showed that IL10/CpG induced proliferation and CDK4 expression were JAK/STAT3 and IKK/NF-κB dependent. This was further supported by STAT3 and p65/RELA knockdown experiments, showing strongest effects on cell proliferation and CDK4 expression after double knockdown. Additionally, chromatin immunoprecipitation revealed a dual binding of STAT3 and p65 to the proximal promotor of CDK4 after IL10/CpG treatment. Therefore, the observed synergism of IL10R and TLR9 signalling was able to induce proliferation in a comparable way as aberrant MYC and might play a role in B cell homeostasis or transformation.
Clinic of Haematology and Medical Oncology, University Medical Centre of the Georg-August University Göttingen, Göttingen, Germany.|BMBF e:Bio Network, MMML-Myc-Sys.
Maren
M
Feist
Clinic of Haematology and Medical Oncology, University Medical Centre of the Georg-August University Göttingen, Göttingen, Germany.
Judith
J
Kemper
BMBF e:Bio Network, MMML-Myc-Sys.|Statistical Bioinformatics, Institute of Functional Genomics, University of Regensburg, Germany.
Franziska
F
Taruttis
Statistical Bioinformatics, Institute of Functional Genomics, University of Regensburg, Germany.
Thorsten
T
Rehberg
BMBF e:Bio Network, MMML-Myc-Sys.|Statistical Bioinformatics, Institute of Functional Genomics, University of Regensburg, Germany.
Julia C
JC
Engelmann
BMBF e:Bio Network, MMML-Myc-Sys.|Institute of Functional Genomics, University of Regensburg, Germany.
Wolfram
W
Gronwald
BMBF e:Bio Network, MMML-Myc-Sys.|Institute for Pathology, Campus Benjamin Franklin, Charité Berlin, Germany.
Michael
M
Hummel
BMBF e:Bio Network, MMML-Myc-Sys.|Statistical Bioinformatics, Institute of Functional Genomics, University of Regensburg, Germany.
Rainer
R
Spang
Clinic of Haematology and Medical Oncology, University Medical Centre of the Georg-August University Göttingen, Göttingen, Germany.|BMBF e:Bio Network, MMML-Myc-Sys.
Dieter
D
Kube
D001402:B-Lymphocytes / Q000166:cytology / Q000187:drug effects*; D002455:Cell Division; D002461:Cell Line, Transformed; D002471:Cell Transformation, Neoplastic; D002478:Cells, Cultured; D018899:CpG Islands; D051358:Cyclin-Dependent Kinase 4 / Q000037:antagonists & inhibitors / Q000502:physiology; D051361:Cyclin-Dependent Kinase 6 / Q000037:antagonists & inhibitors / Q000502:physiology; D004357:Drug Synergism; D005786:Gene Expression Regulation; D006801:Humans; D016753:Interleukin-10 / Q000494:pharmacology / Q000502:physiology*; D008223:Lymphoma / Q000209:etiology; D016271:Proto-Oncogene Proteins c-myc / Q000502:physiology; D016196:S Phase / Q000187:drug effects; D050796:STAT3 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D051217:Toll-Like Receptor 9 / Q000819:agonists / Q000502:physiology*; D051996:Transcription Factor RelA / Q000378:metabolism
C508609:IL10 protein, human; C489427:MYC protein, human; D016271:Proto-Oncogene Proteins c-myc; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; C495371:TLR9 protein, human; D051217:Toll-Like Receptor 9; D051996:Transcription Factor RelA; D016753:Interleukin-10; C495900:CDK4 protein, human; C495911:CDK6 protein, human; D051358:Cyclin-Dependent Kinase 4; D051361:Cyclin-Dependent Kinase 6
B cell proliferation; cyclin-dependent kinase; interleukin; toll-like receptor