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Assessment of Be1 and Be2 cells in systemic lupus erythematosus indicates elevated interleukin-10 producing CD5+ B cells

Division of Molecular and Clinical Immunology, School of Clinical Laboratory Sciences, University of Nottingham, Queen’s Medical Centre, Nottingham, UK

M L Huggins

Division of Molecular and Clinical Immunology, School of Clinical Laboratory Sciences, University of Nottingham, Queen’s Medical Centre, Nottingham, UK

P Lanyon

Division of Molecular and Clinical Immunology, School of Clinical Laboratory Sciences, University of Nottingham, Queen’s Medical Centre, Nottingham, UK

A Robins

Division of Molecular and Clinical Immunology, School of Clinical Laboratory Sciences, University of Nottingham, Queen’s Medical Centre, Nottingham, UK

R J Powell

Division of Molecular and Clinical Immunology, School of Clinical Laboratory Sciences, University of Nottingham, Queen’s Medical Centre, Nottingham, UK

I Todd

Division of Molecular and Clinical Immunology, School of Clinical Laboratory Sciences, University of Nottingham, Queen’s Medical Centre, Nottingham, UK, ian.todd{at}nottingham.ac.uk

Recent studies indicate that normal B cells can be primed to differentiate into two distinct cytokine- secreting effector subsets, Be1 and Be2. The aim of this study was to analyse, for the first time, Be1 and Be2 cells at the single cell level in SLE patients using the recently developed technique of flow cytometry for intracellular cytokines. Peripheral blood mononuclear cells (PBMC) from SLE patients and age- and sex-matched normal controls were cultured for 24 h in the presence or absence of phorbal myristate acetate and ionomycin (PMA=I) or lipopolysaccharide (LPS). The production of type 1 (IFN-g, IL-2) and type 2 (IL-4, IL-5, IL-6, IL-10, IL-13) cytokines by B cells (and IL-10 production by fractionated CD5⁺ and CD5⁷ B cells) was investigated using an intracellular cytokine staining technique and flow cytometry. In the absence of PMA=I stimulation, the percentage of B cells from SLE patients was significantly lower than those of normal subjects and significantly more SLE B cells spontaneously produced IL-10 than controls. Moreover, CD5⁺ B cells from SLE patients were enriched for cells with signs of previous in vivo activation and for high levels of IL-10 production. A significant positive correlation was observed between the percentage of IL-10- and IL-6-producing PMA=I-stimulated B cells in SLE patients, but not in controls. There were no significant differences in the production of other cytokines by B cells of SLE patients and normal subjects. In conclusion, a general alteration of type 1 and type 2 cytokine production by B cells is not observed in SLE patients. The role of B cell cytokines in the pathogenesis of SLE appears to be exerted by elevated secretion of in vivo IL-10, which may play an important role in the immune dysregulation observed in SLE patients. Moreover, the cross regulation of IL-10 and IL-6 is disrupted in SLE patients.

Key Words: Be1 cells • Be2 cells • CD5+ B cells • cytokines • systemic lupus erythematosus

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