title: IL-10 production by CLL cells is enhanced in the anergic IGHV mutated subset and associates with reduced DNA methylation of the IL10 locus.
abstract: Chronic lymphocytic leukemias (CLLs) with unmutated (U-CLL) or mutated (M-CLL) IGHV have variable features of immunosuppression, possibly influenced by those CLL cells activated to produce interleukin 10 (IL-10). The two subsets differ in their levels of anergy, defined by low surface immunoglobulin M levels/signaling capacity, and in their DNA methylation profile, particularly variable in M-CLL. We have now found that levels of IL-10 produced by activated CLL cells were highly variable. Levels were higher in M-CLL than in U-CLL and correlated with anergy. DNA methylation analysis of IL10 locus revealed two previously uncharacterized 'variably methylated regions' (CLL-VMRs1/2) in the gene body, but similarly low methylation in the promoter of both U-CLL and M-CLL. CLL-VMR1/2 methylation was lower in M-CLL than in U-CLL and inversely correlated with IL-10 induction. A functional signal transducer and activator of transcription 3 (STAT3) binding site in CLL-VMR2 was confirmed by proximity ligation and luciferase assays, whereas inhibition of SYK-mediated STAT3 activation resulted in suppression of IL10. The data suggest epigenetic control of IL-10 production. Higher tumor load may compensate the reduced IL-10 production in U-CLL, accounting for clinical immunosuppression in both subsets. The observation that SYK inhibition also suppresses IL-10 provides a potential new rationale for therapeutic targeting and immunological rescue by SYK inhibitors in CLL.
authors.affiliation: Haematology Oncology Group, Cancer Sciences Unit, Cancer Research UK and NIHR Experimental Cancer Medicine Centres, Faculty of Medicine, University of Southampton, Southampton, UK.
authors.forename: S
authors.initials: S
authors.lastname: Drennan
authors.affiliation: Haematology Oncology Group, Cancer Sciences Unit, Cancer Research UK and NIHR Experimental Cancer Medicine Centres, Faculty of Medicine, University of Southampton, Southampton, UK.
authors.forename: A
authors.initials: A
authors.lastname: D'Avola
authors.affiliation: Wessex Investigational Sciences Hub laboratory, Cancer Sciences Unit, Cancer Research UK and NIHR Experimental Cancer Medicine Centres, Faculty of Medicine, University of Southampton, Southampton, UK.
authors.forename: Y
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Division of Epigenomics and Cancer Risk Factors, The German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Weigel
authors.affiliation: Haematology Oncology Group, Cancer Sciences Unit, Cancer Research UK and NIHR Experimental Cancer Medicine Centres, Faculty of Medicine, University of Southampton, Southampton, UK.
authors.forename: E
authors.initials: E
authors.lastname: Chrysostomou
authors.affiliation: Molecular Oncology Group, Cancer Sciences Unit, Cancer Research UK and NIHR Experimental Cancer Medicine Centres, Faculty of Medicine, University of Southampton, Southampton, UK.
authors.forename: A J
authors.initials: AJ
authors.lastname: Steele
authors.affiliation: Department of Translational Oncology, National Center for Tumor Diseases (NCT) and German Cancer Research Center (DKFZ), Heidelberg, Germany.|Department of Medicine V, University of Heidelberg, Heidelberg, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Zenz
authors.affiliation: Division of Epigenomics and Cancer Risk Factors, The German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: C
authors.identifier: 0000-0003-2554-3952
authors.initials: C
authors.lastname: Plass
authors.affiliation: Medical Oncology, University Hospital Southampton National Health Service Trust, Southampton, UK.
authors.forename: P W
authors.initials: PW
authors.lastname: Johnson
authors.affiliation: Wessex Investigational Sciences Hub laboratory, Cancer Sciences Unit, Cancer Research UK and NIHR Experimental Cancer Medicine Centres, Faculty of Medicine, University of Southampton, Southampton, UK.
authors.forename: A P
authors.initials: AP
authors.lastname: Williams
authors.affiliation: Molecular Oncology Group, Cancer Sciences Unit, Cancer Research UK and NIHR Experimental Cancer Medicine Centres, Faculty of Medicine, University of Southampton, Southampton, UK.
authors.forename: G
authors.initials: G
authors.lastname: Packham
authors.affiliation: Molecular Immunology Group, Cancer Sciences Unit, Cancer Research UK and NIHR Experimental Cancer Medicine Centres, Faculty of Medicine, University of Southampton, Southampton, UK.
authors.forename: F K
authors.initials: FK
authors.lastname: Stevenson
authors.affiliation: Division of Epigenomics and Cancer Risk Factors, The German Cancer Research Center (DKFZ), Heidelberg, Germany.|Division of Hematology, The Ohio State University, Columbus, OH, USA.
authors.forename: C C
authors.initials: CC
authors.lastname: Oakes
authors.affiliation: Haematology Oncology Group, Cancer Sciences Unit, Cancer Research UK and NIHR Experimental Cancer Medicine Centres, Faculty of Medicine, University of Southampton, Southampton, UK.|Haematology Department, University Hospital Southampton National Health Service Trust, Southampton, UK.
authors.forename: F
authors.identifier: 0000-0002-2211-1831
authors.initials: F
authors.lastname: Forconi
mesh_terms: D019175:DNA Methylation; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D016753:Interleukin-10 / Q000096:biosynthesis* / Q000235:genetics; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000276:immunology*; D009154:Mutation; D050796:STAT3 Transcription Factor / Q000378:metabolism; D000072377:Syk Kinase / Q000037:antagonists & inhibitors / Q000502:physiology
chemical_list: C508609:IL10 protein, human; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D016753:Interleukin-10; D000072377:Syk Kinase