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

Abstract

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.

Keywords

Be1 cells Be2 cells CD5+ B cells cytokines systemic lupus erythematosus

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References

Romagnani S. Human TH1 and TH2 subsets: doubt no more . Immunol Today 1991; 12: 256-257 .

Harris DP , Haynes L , Sayles PC , Duso DK , Eaton SM , Lepak NM , Johnson LL , Swain SL , Lund FE. Reciprocal regulation of polarized cytokine production by effector B and T cells . Nat Immunol 2000; 1: 475-481 .

Stollar B. The antigenic potential and specificity of nucleic acids, nucleoproteins, and their modified derivatives . Arthritis Rheum 1981; 28: 1010-1017 .

Liossis SC , Tsokos GC. B cells in systemic lupus erythematosus. In: Kammer GM , Tosoko GC (eds). Lupus: Molecular and Cellular Pathogenesis. Humana Press: Totowa, NJ , 1999, pp 167-180.

Cross JT , Benton HP. The role of interleukin-6 and interleukin-10 in B cell hyperactivity in systemic lupus erythematosus . inflamm Res 1999; 48: 255-261 .

Amel Kashipaz MR , Huggins ML , Lanyon P , Todd I , Powell RJ. Quantitative and qualitative analysis of balance between type 1 and type 2 cytokine-producing CD8+ and CD87 T cells in systemic lupus erythematosus . J Autoimmun 2001; 17: 155-163 .

Tan EM , Cohen AS , Fries JF et al. The 1982 revised ceiteria for the classification of systemic lupus erythematosus . Arthritis Rheum 1982; 25: 1271-1277 .

Howard M , O’Garra A. Biological properties of interleukin 10 . Immunol Today 1992; 13: 198-200 .

de Waal Melefyt R , Yssel H , de Vries JE. Direct effect of IL-10 on subsets of human CD4+ T cells clones and resting T cells. Specific inhibition of IL-2 production and proliferation . J Immunol 1993; 150: 4754-4765 .

10.

Georgescu L , Krishna Vakkalanka R , Elkon KB , Crow MK. Interleukin-10 promotes activation-induced cell death of SLE lymphocytes mediated by Fas ligand . J Clin Invest 1997; 100: 2622-2633 .

11.

Schmidt M , Lugering N , Pauels HG , Schulze-Osthoff K , Domschke W , Kucharzik T. IL-10 induces apoptosis in human monocytes involving the CD95 receptor/ligand pathway . Eur J Immunol 2000; 30: 1769-1777 .

12.

Bruns A , Blass S , Hausdorf G , Burmester GR , Hiepe F. Nucleosomes are major T and B cell autoantigens in systemic lupus erythematosus . Arthritis Rheum 2000; 43: 2307-2315 .

13.

Voll RE , Herrmann M , Roth EA , Stach C , Kalden JR , Girkontaite I. Immunosuppressive effects of apoptotic cells . Nature 1997; 390: 350-351 .

14.

Pers JO , Jamin C , Predine-Hug F , Lydyard P , Youinou P. The role of CD57 expressing B cells in health and disease . Int J Mol Med 1999; 3: 239-245 .

15.

O’Garra A , Chang R , Haughton G , Howard M. Ly-1 B (B1) cells are the main source of B cell-derived interleukin 10 . Eur J Immunol 1992; 22: 711-717 .

16.

Bhat NM , Kantor AB , Bieber MM , Stall AM , Herzenberg LA , Teng NN. The ontogeny and functional characteristics of human B-1 (CD5+) B cells . Int Immunol 1992; 4: 243-252 .

17.

Nisitani S , Murakami M , Akamizu T , Okino T , Ohmori K , Mori T , Imamura M , Honjo T. Preferential localization of human cd5(+) B cells in the peritoneal cavity . Scand J Immunol 1997; 46: 541-545 .

18.

Casali P , Notkins AL. CD5+ B lymphocytes, polyreactive antibodies and the human B-cell repertoire . Immunol Today 1989; 10: 364-368 .

19.

Klinman DM , Shirai A , Conover J , Steinberg AD. Cross-reactivity of IgG anti-DNA-secreting B cells in patients with systemic lupus erythematosus . Eur J Immunol 1994; 24: 53-58 .

20.

Werner-Favre C , Vischer TL , Wohlwend D , Zubler RH. Cell surface antigen CD5 is a marker for activated human B cells . Eur J Immunol 1989; 19: 1209-1213 .

21.

Moore KW , O’Garra A , de Waal Maleyft R. Interleukin-10 . A Rev Immunol 1993; 11: 165-190 .

22.

Mongan AE , Ramdahin S , Warrington RJ. IL-10 response abnormalities in systemic lupus erythematosus . Scand J Immunol 1997; 46: 406-412 .

23.

Klinman DM , Steinberg AD. Inquiry into murine and human lupus . Immunol Rev 1995; 144: 157-193 .

24.

Levy Y , Brouet JC. Interleukine-10 prevents spontaneous death of germinal center B cells by induction of the bcl-2 protein . J Clin Invest 1994; 93: 424-428 .

25.

Itoh K , Hirohata S. The role of IL-10 in human B cell activation, proliferation, and differentiation . J Immunol 1995; 154: 4341-4350 .

26.

Li L , Krajewski S , Reed JC , Choi YS. The apoptosis and proliferation of SAC-activated B cells by IL-10 are associated with changes in Bcl-2, Bcl-xL, and Mcl-1 expression . Cell Immunol 1997; 178: 33-41 .

27.

Segal R , Dayan M , Zinger H , Mozes E. Methotrexate treatment in murine experimental systemic lupus erythematosus (SLE); clinical benefits associated with cytokine manipulation . Clin Exp Immunol 1995; 101: 66-72 .

28.

Ishida H , Muchamuel T , Sakaguchi S , Andrade S , Menon S , Howard M. Continuousadministration of anti-interleukin 10 antibodies delays onset of autoimmunity in NZB=W F1 mice . J Exp Med 1994; 179: 305-310 .

29.

Llorente L , Richaud-Patin Y , Garcia-Padilla C et al. Clinical and biological effects of anti-IL-10 monoclonal antibody administration in systemic lupus erythematosus . Arthritis Rheum 2000; 43: 1790-1800 .

30.

Prud’homme GJ. Gene therapy of autoimmune diseases with vectors encoding regulatory cytokines or inflammatory cytokine inhibitors . J Gene Med 2000; 2: 222-232 .