B-lymphocyte subpopulations in patients with rheumatoid arthritis and the effect of an interleukin-6 receptor inhibitor on them.

The clinical efficacy and safety of interleukin-6 (IL-6) receptor blockade have been well studied, but the data on the impact of therapeutic inhibition of IL-6 on B cells are scarce and contradictory. Preliminary reports have shown that B cell function and a humoral immune response may be modulated by an IL-6 receptor inhibitor.
Objective: to assess the effect of 12-month tocilizumab (TCZ) therapy on B-cell phenotype and gene expression in RA and to analyze the association between B-cell subsets and RA activity.
Subjects and methods. Examinations were made in 24 active RA patients (20 women and 4 men) (median age, 55 [49; 64] years; disease duration, 72 [24; 108] months; DAS28 5.8 [5.3; 6.3]; the patients were seropositive for rheumatoid factor (RF) (100%) and for anti-cyclic citrullinated peptide antibodies (87.3%). The patients received TCZ 8 mg/kg every 4 weeks. After 12 months of therapy, 54% of patients were categorized as good responders, 46% as moderate responders according to the EULAR response criteria. A control group consisted of 29 volunteers (21 women and 8 men; median age, 58.5 [53.0; 62.0] years). Peripheral blood lymphocytes were immunophenotyped at the time of enrollment and after 12 months. The absolute and relative counts of CD19+B lymphocytes, memory B cells (CD19+CD27+), non-switched memory B cells (CD19+IgD+CD27+), switched memory B cells (CD19+IgDCD27+), naive (CD19+IgD+CD27-), double-negative (CD19+IgD-CD27-), transitional (CD19+IgD+CD10+CD38++CD27) B cells, plasma cells (CD19+СD38+), and plasmablasts (CD19+СD38+++IgD-CD27+CD20-) were estimated using multicolor flow cytometry.
Results and discussion. The relative and absolute counts of memory B cells (CD19+CD27+) (1.3 [0.9; 1.7]%, 0015 [0.001; 0.003]•109/l), switched memory B cells (CD19+IgD-CD27+) (6.8 [3.6; 11.6]%, 0.01 [0.005; 0.02]•109/l), and the absolute number of transitional B cells (CD19+CD38++CD10+IgD+CD27-) (0.00009 [0; 0.00028]•109/l) were found to be lower in RA patients than in donors: 2.2 [1.1; 3.0]%, 0.003 [0.001; 0.007]•109/l; 12.8 [9.3; 17.0]%, 0.02 [0.01; 0.04]•109/l; 0.0001 [0; 0.0003]•109/l, respectively (p<0.05 for all cases). After 12 months of TCZ therapy initiation, there were decreases in the relative and absolute counts of plasmablasts (CD19+CD38+++CD27+IgD-CD20-) from 0.15 [0.1; 0.3] to 0.1 [0.01; 0.1]% and from 0.0003 [0.00007; 0.004]•109/l to 0.0001 [0; 0.0003]•109/l, respectively (p<0.05). At the same time, the relative and absolute counts of memory B cells (CD19+CD27+) and switched memory B cells (CD19+CD27+IgD-) remained lower in RA patients than in donors: 1.0 [0.7; 1.2] and 2.2 [1.1; 3.0]%; 0.001 [0.006; 0.003]•109/l and 0.003 [0.001; 0.007]•109/l; 3.1 [1.1; 4.2] and 12.8 [9.3; 17.0]%; 0.003 [0.002; 0.006]•109/l and 0.02 [0.01; 0.04]•109/l, respectively (p<0.05 for all cases). Following 12 months of TCZ therapy, the numbers of other B-cell subpopulations were not considerably altered. When included in the study, the patients with RA showed correlations between the absolute count of memory B cells (CD19+CD27+) and the level of C-reactive protein (r=0.50; p<0.05); between the absolute count of plasmablasts (CD19+CD38+++CD27+IgD-CD20-) and the level of RF (r=0.41 and r=0.52; p<0.05). There were no correlations of B cell subsets with clinical and laboratory findings after 12 months of TCZ initiation.
Conclusion. Immunophenotyping of peripheral blood B lymphocyte subsets showed the lower relative and absolute counts of memory B cells (CD19+CD27+) and switched memory B cells (CD19+CD27+IgD-) in RA patients than in healthy donors. The found correlations between the counts of memory B cells and plasmablasts and the values of laboratory parameters in patients with high RA activity may suggest that B lymphocytes are involved in the pathogenesis of RA. There was a decline in plasmablast levels after 12 months of TCZ therapy.

1. Насонов ЕЛ. Проблемы иммунопатологии ревматоидного артрита: эволюция болезни. Научно-практическая ревматология. 2017;55(3):277-94 [Nasonov EL. Рroblems of rheumatoid arthritis immunopathology: evolution of the disease. Nauchno-Prakticheskaya Revmatologiya = Rheumatology Science and Practice. 2017;55(3):277-94 (In Russ.)]. doi: 10.14412/1995-4484-2017-277-294

2. Leslie D, Lipsky P, Notkins AL. Autoantibodies as predictors of disease. J Clin Invest. 2001;108:1417-22. doi: 10.1172/JCI14452

3. Leandro M. B cells and rheumatoid factors in autoimmunity. Int J Rheum Dis. 2015;18:379-81. doi: 10.1111/1756-185X.12690

4. Lino AC, Dorner T, Bar-Or A, Fillatreau S. Cytokine-producing B cells: a translational view on their roles in human and mouse autoimmune diseases. Immunol Rev. 2016;269:130-44. doi: 10.1111/imr.12374

5. Насонов ЕЛ, Лила АМ. Ингибиция интерлейкина 6 при иммуновоспалительных ревматических заболеваниях: достижения, перспективы и надежды. Научно-практическая ревматология. 2017;55(6):590-9 [Nasonov EL, Lila AM. Inhibition of interleukin 6 in immune inflammatory rheumatic diseases: achievements, prospects, and hopes. Nauchno-Prakticheskaya Revmatologiya = Rheumatology Science and Practice. 2017;55(6):590-9 (In Russ.)]. doi: 10.14412/1995-4484-2017-590-599

6. Scott LJ. Tocilizumab: A Review in Rheumatoid Arthritis. Drugs. 2017 Nov;77(17):1865-79. doi: 10.1007/s40265-017-0829-7

7. Yeo L, Toellner KM, Salmon M, et al. Cytokine mRNA profiling identifies B cells as a major source of RANKL in rheumatoid arthritis. Ann Rheum Dis. 2011 Nov;70(11):2022-8. doi: 10.1136/ard.2011.153312

8. Mihara M, Hashizume M, Yoshida H, et al. IL-6/IL-6 receptor system and its role in physiological and pathological conditions. Clin Sci (Lond). 2012 Feb;122(4):143-59. doi: 10.1042/CS20110340

9. Burmester GR, Feist E, Dö rner T. Emerging cell and cytokine targets in rheumatoid arthritis. Nat Rev Rheumatol. 2014 Feb;10(2):77-88. doi: 10.1038/nrrheum.2013.168

10. Roll P, Muhammad K, Schumann M, et al. In vivo effects of the anti-interleukin-6 receptor inhibitor tocilizumab on the B cell compartment. Arthritis Rheum. 2011 May;63(5):1255-64. doi: 10.1002/art.30242

11. Moura RA, Quaresma C, Vieira AR, et al. B-cell phenotype and IgD-CD27- memory B cells are affected by TNF-inhibitors and tocilizumab treatment in rheumatoid arthritis. PLoS One. 2017 Sep 8;12(9):e0182927. doi: 10.1371/journal.pone.0182927

12. Lubbers J, Vosslamber S, van de Stadt LA, et al. B cell signature contributes to the prediction of RA development in patients with arthralgia. Ann Rheum Dis. 2015;74:1786-8. doi: 10.1136/annrheumdis-2015-207324

13. Van Baarsen LGM, de Hair MJH, Ramwadhdoebe TH, et al. The cellular composition of lymph nodes in the earliest phase of inflammatory arthritis. Ann Rheum Dis. 2013;72:1420-4. doi: 10.1136/annrheumdis-2012-202990

14. Amara K, Steen J, Murray F, et al. Monoclonal IgG antibodies generated from joint-derived B cells of RA patients have a strong bias toward citrullinated autoantigen recognition. J Exp Med. 2013;210:445-55. doi: 10.1084/jem.20121486

15. Li S, Yu Y, Yue Y, et al. Autoantibodies from single circulating plasmablasts react with citrullinated antigens and Porphyromonas gingivalis in rheumatoid arthritis. Arthritis Rheum. 2016;68:614-26. doi: 10.1002/art.39455

16. Muhammad K, Roll P, Einsele H, et al. Delayed acquisition of somatic hypermutations in repopulated IGD+CD27+ memory B cell receptors after rituximab treatment. Arthritis Rheum. 2009;60:2284-93. doi: 10.1002/art.24722

17. Adlowitz DG, Barnard J, Biear J, et al. Expansion of Activated Peripheral Blood Memory B Cells in Rheumatoid Arthritis, Impact of B Cell Depletion Therapy, and Biomarkers of Response. PLoS One. 2015 Jun 5;10(6):e0128269. doi: 10.1371/journal.pone.0128269

18. Fekete A, Soos L, Szekanecz Z, et al. Disturbances in B- and T-cell homeostasis in rheumatoid arthritis: suggested relationships with antigen-driven immune responses. J Autoimmun. 2007;29:154-63. doi: 10.1016/j.jaut.2007.07.002

19. Moura RA, Graca L, Fonseca JE. To B or not to B the conductor of rheumatoid arthritis orchestra. Clin Rev Allergy Immunol. 2012 Dec;43(3):281-91. doi: 10.1007/s12016-012-8318-y

20. Супоницкая ЕВ, Алексанкин АП, Александрова ЕН и др. Определение субпопуляций В-лимфоцитов периферической крови методом проточной цитофлуорометрии у здоровых лиц и больных ревматическими заболеваниями. Клиническая лабораторная диагностика. 2015;60(6):30-33 [Suponitskaya EV, Aleksankin AP, Alexandrova EN, et al. Characterization of peripheral blood b-cell subset in patients with systemic lupus erythematosus. Klinicheskaya Laboratornaya Diagnostika. 2015;60(6):30-3 (In Russ.)].

21. Nakayamada S, Kubo S, Yoshikawa M, et al. Differential effects of biological DMARDs on peripheral immune cell phenotypes in patients with rheumatoid arthritis. Rheumatology (Oxford). 2018 Jan 1;57(1):164-74. doi: 10.1093/rheumatology/kex012

22. Lü bbers J, van Beers-Tas MH, Vosslamber S, et al. Changes in peripheral blood lymphocyte subsets during arthritis development in arthralgia patients. Arthritis Res Ther. 2016;18(1):205. doi: 10.1186/s13075-016-1102-2

23. Moura RA, Weinmann P, Pereira PA, et al. Alterations on peripheral blood B-cell subpopulations in very early arthritis patients. Rheumatology (Oxford). 2010;49:1082-92. doi: 10.1093/rheumatology/keq029

24. Mahmood Z, Muhammad K, Schmalzing M, et al. CD27-IgDmemory B cells are modulated by in vivo interleukin-6 receptor (IL-6R) blockade in rheumatoid arthritis. Arthritis Res Ther. 2015 Mar 14;17:61. doi: 10.1186/s13075-015-0580-y

25. Illei GG, Shirota Y, Yarboro CH, et al. Tocilizumab in systemic lupus erythematosus: data on safety, preliminary efficacy, and impact on circulating plasma cells from an open-label phase I dosage-escalation study. Arthritis Rheum. 2010;62:542-52. doi: 10.1002/art.27221

26. Меснянкина АА, Соловьев СК, Александрова ЕН и др. Динамика субпопуляции В-лимфоцитов у больных системной красной волчанкой на фоне терапии генно-инженерными биологическими препаратами. Научно-практическая ревматология. 2017;55(3):252-60 [Mesnyankina AA, Solovyev SK, Aleksandrova EN, et al. The time course of changes in B lymphocyte subpopulations in patients with systemic lupus erythematosus during therapy with biological agents. Nauchno-Prakticheskaya Revmatologiya = Rheumatology Science and Practice. 2017;55(3):252-60 (In Russ.)]. doi: 10.14412/1995-4484-2017-252-260

27. Marie-Cardine A, Divay F, Dutot I, et al. Transitional B cells in humans: characterization and insight from B lymphocyte reconstitution after hematopoietic stem cell transplantation. Clin Immunol. 2008;127:14-25. doi: 10.1016/j.clim.2007.11.013

28. Fonseca JE, Santos MJ, Canhao H, Choy E. Interleukin-6 as a key player in systemic inflammation and joint destruction. Autoimmun Rev. 2009;8:538-42. doi: 10.1016/j.autrev.2009.01.012

29. Jourdan M, Cren M, Robert N, et al. IL-6 supports the generation of human long-lived plasma cells in combination with either APRIL or stromal cell-soluble factors. Leukemia. 2014 Aug;28(8):1647-56. doi: 10.1038/leu.2014.61

30. Muhammad K, Roll P, Seibold T, et al. Impact of IL-6 receptor inhibition on human memory B cells in vivo: impaired somatic hypermutation in preswitch memory B cells and modulation of mutational targeting in memory B cells. Ann Rheum Dis. 2011;70:1507-10. doi: 10.1136/ard.2010.141325

31. Kikuchi J, Hashizume M, Kaneko Y, et al. Peripheral blood CD4(+)CD25(+)CD127(low) regulatory T cells are significantly increased by tocilizumab treatment in patients with rheumatoid arthritis: increase in regulatory T cells correlates with clinical response. Arthritis Res Ther. 2015 Jan 21;17:10. doi: 10.1186/s13075-015-0526-4

32. Fleischer S, Ries S, Shen P, et al. Anti-interleukin-6 signalling therapy rebalances the disrupted cytokine production of B cells from patients with active rheumatoid arthritis. Eur J Immunol. 2018 Jan;48(1):194-203. doi: 10.1002/eji.201747191

33. Alivernini S, Kurowska-Stolarska M, Tolusso B, et al. MicroRNA-155 influences B-cell function through PU.1 in rheumatoid arthritis. Nat Commun. 2016 Sep 27;7:12970. doi: 10.1038/ncomms12970

34. Wu Y, El Shikh ME, El Sayed RM, et al. IL-6 produced by immune complex-activated follicular dendritic cells promotes germinal center reactions, IgG responses and somatic hypermutation. Int Immunol. 2009;21:745-56. doi: 10.1093/intimm/dxp041