title: TLR9 mediated regulatory B10 cell amplification following sub-total body irradiation: Implications in attenuating EAE.
abstract: Autoimmunity and inflammation are controlled in part by regulatory B (Breg) cells, including the recently identified IL-10-competent B10 cell subset that represents 1%-3% of mouse spleen B cells. In this study, the influence of irradiation on Breg/B10 cell generation and IL-10 production mediated by TLR9 signaling pathways was investigated. Spleen and peritoneal cavity Breg/B10 cell frequencies were significantly expanded three weeks after sub-total body irradiation (sub-TBI, 5Gy or 10Gy) in adult male wild type (WT) C57BL/6(B6) mice but not in TLR9-/-mice. TLR9 agonist ODN1826 stimulation in vitro for 5h induced more B10 cells to express cytoplasmic IL-10 in sub-TBI WT mice than in TLR9-/-mice. Prolonged ODN1826 stimulation (48h) induced additional spleen CD19hiCD5+CD1dhiB cells to express IL-10. TLR9-dependent signaling molecules, MyD88, TRAF6 and IRF8 are involved in sub-TBI induced Breg/B10 cells development and expansion. Furthermore, using a mouse model for multiple sclerosis, we show here that sub-TBI induced Breg/B10 cells dramatically inhibit disease onset and severity when transferred into mice with established experimental autoimmune encephalomyelitis (EAE). Adoptively transferred sub-TBI induced Breg cells significantly suppress inflammatory T cell responses of TH17 and TH1 types in EAE mice. In conclusion, sub-TBI can drive Breg/B10 cell development and expansion, which could be used as a novel tool for suppressing undesirable immunity. The ex vivo expansion and reinfusion of autologous Breg/B10 cells may provide a novel and effective in vivo treatment for severe autoimmune diseases that are resistant to current therapies.
authors.affiliation: Central Lab., First Affiliated Hospital, Fujian Medical University, Fujian Key Laboratory of Individualized Active Immunotherapy, Fujian Medical University, Key Laboratory of Radiation Biology (Fujian Medical University), Fujian Province University, Fuzhou, Fujian 350005, China; Department of Radiation Oncology, First Affiliated Hospital, Fujian Medical University, Fuzhou, Fujian 350005, China.
authors.forename: Jinsheng
authors.initials: J
authors.lastname: Hong
authors.affiliation: Department of Dermatology, Yangpu Hospital, Tongji University School of Medicine, Shanghai 200090, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Fang
authors.affiliation: Central Lab., First Affiliated Hospital, Fujian Medical University, Fujian Key Laboratory of Individualized Active Immunotherapy, Fujian Medical University, Key Laboratory of Radiation Biology (Fujian Medical University), Fujian Province University, Fuzhou, Fujian 350005, China; Department of Radiation Oncology, First Affiliated Hospital, Fujian Medical University, Fuzhou, Fujian 350005, China.
authors.forename: Ruilong
authors.initials: R
authors.lastname: Lan
authors.affiliation: National Key Laboratory of Medical Immunology & Institute of Immunology, Second Military Medical University, Shanghai 200433, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Tan
authors.affiliation: National Key Laboratory of Medical Immunology & Institute of Immunology, Second Military Medical University, Shanghai 200433, China.
authors.forename: Yeping
authors.initials: Y
authors.lastname: Tian
authors.affiliation: Department of Radiation Oncology, UF Shands Cancer Center, Gainesville, FL, United States.
authors.forename: Mei
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Department of Radiation Oncology, UF Shands Cancer Center, Gainesville, FL, United States.
authors.forename: Paul
authors.initials: P
authors.lastname: Okunieff
authors.affiliation: Central Lab., First Affiliated Hospital, Fujian Medical University, Fujian Key Laboratory of Individualized Active Immunotherapy, Fujian Medical University, Key Laboratory of Radiation Biology (Fujian Medical University), Fujian Province University, Fuzhou, Fujian 350005, China; Department of Radiation Oncology, First Affiliated Hospital, Fujian Medical University, Fuzhou, Fujian 350005, China.
authors.forename: Lurong
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Central Lab., First Affiliated Hospital, Fujian Medical University, Fujian Key Laboratory of Individualized Active Immunotherapy, Fujian Medical University, Key Laboratory of Radiation Biology (Fujian Medical University), Fujian Province University, Fuzhou, Fujian 350005, China; Department of Radiation Oncology, First Affiliated Hospital, Fujian Medical University, Fuzhou, Fujian 350005, China.
authors.forename: Jianhua
authors.initials: J
authors.lastname: Lin
authors.affiliation: Central Lab., First Affiliated Hospital, Fujian Medical University, Fujian Key Laboratory of Individualized Active Immunotherapy, Fujian Medical University, Key Laboratory of Radiation Biology (Fujian Medical University), Fujian Province University, Fuzhou, Fujian 350005, China; Department of Radiation Oncology, First Affiliated Hospital, Fujian Medical University, Fuzhou, Fujian 350005, China. Electronic address: han18305956512@sina.com.
authors.forename: Deping
authors.initials: D
authors.lastname: Han
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D016175:B-Lymphocyte Subsets / Q000276:immunology / Q000528:radiation effects*; D060151:B-Lymphocytes, Regulatory / Q000276:immunology / Q000528:radiation effects*; D015153:Blotting, Western; D002454:Cell Differentiation / Q000276:immunology / Q000528:radiation effects; D049109:Cell Proliferation / Q000528:radiation effects; D002469:Cell Separation; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D060888:Real-Time Polymerase Chain Reaction; D051217:Toll-Like Receptor 9 / Q000276:immunology*; D014916:Whole-Body Irradiation / Q000379:methods*
chemical_list: C495370:Tlr9 protein, mouse; D051217:Toll-Like Receptor 9
keywords: Autoimmunity; Breg/B10 cells; IL-10; MyD88; TLR9