At present, anti-CD20 monoclonal antibody treatments targeting systemic lupus erythematosus (SLE) are complex, variable, and often have disappointing outcomes. High levels of programmed cell death-1 (PD-1) and its ligands (PD-L1, PD-L2) or CD80/CD86 on B cell surfaces are markers of increased B cell activity. However, their expression levels on CD19+CD20+/− B cells and their clinical significance for SLE dynamics have not been carefully investigated.
Flow cytometry was used to detect the expression levels of PD-1, PD-L1, PD-L2, CD80, and CD86 on CD19+CD20+/− B cells in peripheral blood from SLE patients and healthy controls (HCs). The amount of anti-dsDNA and immunoglobin G (IgG) secreted by CD19+CD20+/− B cells was measured by enzyme-linked immunosorbent assay.
CD19+CD20− B cell frequency was significantly higher in SLE patients than in HCs (P < 0.001), and was positively correlated with disease activity. In SLE patients, frequencies of PD-1, PD-L1, PD-L2, and CD86 on CD19+CD20− B cells were significantly higher than CD19+CD20+ B cells (P ≤ 0.002) and were significantly correlated with individual laboratory and clinically based parameters (P < 0.05). In vitro tests, we found that the levels of anti-dsDNA and IgG secreted by CD19+CD20− B cells from patients with SLE were significantly higher than the HC group (P < 0.05).
We found abnormal frequency of CD19+CD20− B cells and increased expression of surface markers on these cells from SLE patients. And the CD19+CD20− B cells had the ability to proliferate and secrete anti-dsDNA and IgG. Additionally, our results suggested that CD19+CD20− B cells from SLE patients may be the activated B cells and caused poor efficacy of rituximab.
• CD19+CD20−B cell frequencies were significantly higher in SLE patients.
• Frequencies of PD-1 and its ligands on CD19+CD20−B cells increased significantly in SLE patients.
• CD19+CD20−B cells in SLE patients had the ability to secrete anti-dsDNA and IgG.
• CD19+CD20−B cells in SLE patients may be the activated B cells and caused poor efficacy of rituximab.
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carboxy fluorescence in succinimidyl ester
enzyme-linked immunosorbent assay
erythrocyte sedimentation rate
neuropsychiatric systemic lupus erythematosus
peripheral blood mononuclear cells
programmed death 1
programmed death ligand 1
programmed death ligand 2
Statistical Package for Social Sciences
systemic lupus erythematosus
systemic lupus erythematosus disease activity index
follicular helper T-cells
Sang A, Zheng YY, Morel L (2014) Contributions of B cells to lupus pathogenesis. Mol Immunol 62(2):329–338. https://doi.org/10.1016/j.molimm.2013.11.013
Moulton VR, Tsokos GC (2015) T cell signaling abnormalities contribute to aberrant immune cell function and autoimmunity. J Clin Invest 125(6):2220–2227. https://doi.org/10.1172/JCI78087
Santos MAO, Lima MM (2017) CD20 role in pathophysiology of Hodgkin’s disease. Rev Assoc Med Bras 63(9):810–813. https://doi.org/10.1590/1806-9282.63.09.810
Maloney DG (2012) Anti-CD20 antibody therapy for B-cell lymphomas. New l J Med 366(21):2008–2016. https://doi.org/10.1056/NEJMct1114348
Liang Y, Tedder TF (2001) Identification of a CD20-, FcepsilonRIbeta-, and HTm4-related gene family: sixteen new MS4A family members expressed in human and mouse. Genomics 72(2):119–127. https://doi.org/10.1006/geno.2000.6472
Merrill JT, Neuwelt CM, Wallace DJ, Shanahan JC, Latinis KM, Oates JC, Utset TO, Gordon C, Isenberg DA, Hsieh HJ, Zhang D, Brunetta PG (2010) Efficacy and safety of rituximab in moderately-to-severely active systemic lupus erythematosus: the randomized, double-blind, phase II/III systemic lupus erythematosus evaluation of rituximab trial. Arthritis Rheum 62(1):222–233. https://doi.org/10.1002/art.27233
Rovin BH, Furie R, Latinis K, Looney RJ, Fervenza FC, Sanchez-Guerrero J, Maciuca R, Zhang D, Garg JP, Brunetta P, Appel G, LUNAR Investigator Group (2012) Efficacy and safety of rituximab in patients with active proliferative lupus nephritis: the Lupus Nephritis Assessment with Rituximab study. Arthritis Rheum 64(4):1215–1226. https://doi.org/10.1002/art.34359
Chen L, Flies DB (2013) Molecular mechanisms of T cell co-stimulation and co-inhibition. Nat Rev Immunol 13(4):227–242. https://doi.org/10.1038/nri3405
Freeman GJ, Long AJ, Iwai Y, Bourque K, Chernova T, Nishimura H, Fitz LJ, Malenkovich N, Okazaki T, Byrne MC, Horton HF, Fouser L, Carter L, Ling V, Bowman MR, Carreno BM, Collins M, Wood CR, Honjo T (2000) Engagement of the PD-1 immunoinhibitory receptor by a novel B7 family member leads to negative regulation of lymphocyte activation. J Exp Med 192(7):1027–1034
Latchman Y, Wood CR, Chernova T, Chaudhary D, Borde M, Chernova I, Iwai Y, Long AJ, Brown JA, Nunes R, Greenfield EA, Bourque K, Boussiotis VA, Carter LL, Carreno BM, Malenkovich N, Nishimura H, Okazaki T, Honjo T, Sharpe AH, Freeman GJ (2001) PD-L2 is a second ligand for PD-1 and inhibits T cell activation. Nat Immunol 2(3):261–268. https://doi.org/10.1038/85330
Dong H, Zhu G, Tamada K, Chen L (1999) B7-H1, a third member of the B7 family, co-stimulates T-cell proliferation and interleukin-10 secretion. Nat Med 5(12):1365–1369. https://doi.org/10.1038/70932
Nurieva RI, Liu X, Dong C (2009) Yin-Yang of costimulation: crucial controls of immune tolerance and function. Immunol Rev 229(1):88–100. https://doi.org/10.1111/j.1600-065X.2009.00769.x
Okazaki T, Maeda A, Nishimura H, Kurosaki T, Honjo T (2001) PD-1 immunoreceptor inhibits B cell receptor-mediated signaling by recruiting src homology 2-domain-containing tyrosine phosphatase 2 to phosphotyrosine. Proc Natl Acad Sci U S A 98:13866–13871. https://doi.org/10.1073/pnas.231486598
Yamazaki T, Akiba H, Iwai H, Matsuda H, Aoki M, Tanno Y, Shin T, Tsuchiya H, Pardoll DM, Okumura K, Azuma M, Yagita H (2002) Expression of programmed death 1 ligands by murine T cells and APC. J Immunol 169:5538–5545
Thibult ML, Mamessier E, Gertner-Dardenne J, Pastor S, Just-Landi S, Xerri L, Chetaille B, Olive D (2013) PD-1 is a novel regulator of human B-cell activation. Int Immunol 25(2):129–137. https://doi.org/10.1093/intimm/dxs098
Jia XY, Zhu QQ, Wang YY, Lu Y, Li ZJ, Li BQ, Tang J, Wang HT, Song CW, Xie CH, Chen LJ (2019) The role and clinical significance of programmed cell death-ligand 1 expressed on CD19B-cells and subsets in systemic lupus erythematosus. Clin Immunol 198:89–99. https://doi.org/10.1016/j.clim.2018.11.015
McKay JT, Haro MA, Daly CA, Yammani RD, Pang B, Swords WE et al (2017) PD-L2 regulates B-1 cell antibody production against phosphorylcholine through an IL-5-dependent mechanism. J Immunol 199(6):2020–2029. https://doi.org/10.4049/jimmunol.1700555
Lee YH, Woo JH, Choi SJ, Ji JD, Song GG (2009) Association of programmed cell death 1 polymorphism and systemic lupus erythematosus: a meta-analysis. Lupus 18(1):9–15. https://doi.org/10.1177/0961203308093923
Folzenlogen D, Hofer MF, Leung DY, Freed JH, Newell MK (1997) Analysis of CD80 and CD86 expression on peripheral blood B-lymphocytes reveals increased expression of CD86 in lupus patients. Clin Immunol Immunopathol 83(3):199–204
Dolff S, Wilde B, Patschan S, Dürig J, Specker C, Philipp T, Kribben A, Witzke O (2007) Peripheral circulating activated B-cell populations are associated with nephritis and disease activity in patients with systemic lupus erythematosus. Scand J Immunol 66(5):584–590. https://doi.org/10.1111/j.1365-3083.2007.02008.x
Bombardier C, Gladman DD, Urowitz MB, Caron D, Chang CH (1992) Derivation of the SLEDAI. A disease activity index for lupus patients. The Committee on Prognosis Studies in SLE. Arthritis Rheum 35(6):630–640
Petri M, Orbai AM, Alarcon GS, Gordon C, Merrill JT, Fortin PR et al (2012) Derivation and validation of the Systemic Lupus International Collaborating Clinics classification criteria for systemic lupus erythematosus. Arthritis Rheum 64(8):2677–2686. https://doi.org/10.1002/art.34473
Nagy G, Koncz A, Perl A (2005) T- and B-cell abnormalities in systemic lupus erythematosus. Crit Rev Immunol 25(2):123–140
Liao J, Chang C, Wu H, Lu Q (2015) Cell-based therapies for systemic lupus erythematosus. Autoimmun Rev 14(1):43–48. https://doi.org/10.1016/j.autrev.2014.10.001
Alshaiki F, Obaid E, Almuallim A, Taha R, El-Haddad H, Almoallim H (2018) Outcomes of rituximab therapy in refractory lupus: a meta-analysis. Eur J Rheumatol 5(2):118–126. https://doi.org/10.5152/eurjrheum.2018.17096
Forsthuber TG, Cimbora DM, Ratchford JN, Katz E, Stüve O (2018) B cell-based therapies in CNS autoimmunity: differentiating CD19 and CD20 as therapeutic targets. Ther Adv Neurol Disord 11:1756286418761697. https://doi.org/10.1177/1756286418761697
Mei HE, Schmidt S, Dörner T (2012) Rationale of anti-CD19 immunotherapy: an option to target autoreactive plasma cells in autoimmunity. Arthritis Res Ther 14(Suppl. 5):S1. https://doi.org/10.1186/ar3909
Looney RJ, Anolik JH, Campbell D, Felgar RE, Young F, Arend LJ, Sloand JA, Rosenblatt J, Sanz I (2004) B cell depletion as a novel treatment for systemic lupus erythematosus: a phase I/II dose-escalation trial of rituximab. Arthritis Rheum 50(8):2580–2589. https://doi.org/10.1002/art.20430
Leandro MJ, Cambridge G, Ehrenstein MR, Edwards JC (2006) Reconstitution of peripheral blood B cells after depletion with rituximab in patients with rheumatoid arthritis. Arthritis Rheum 54(2):613–620. https://doi.org/10.1002/art.21617
Albert D, Dunham J, Khan S, Stansberry J, Kolasinski S, Tsai D, Pullman-Mooar S, Barnack F, Striebich C, Looney RJ, Prak ETL, Kimberly R, Zhang Y, Eisenberg R (2008) Variability in the biological response to anti-CD20 B cell depletion in systemic lupus erythaematosus [J]. Ann Rheum Dis 67(12):1724–1731. https://doi.org/10.1136/ard.2007.083162
Häusler D, Häusser-Kinzel S, Feldmann L, Torke S, Lepennetier G, Bernard CCA, Zamvil SS, Brück W, Lehmann-Horn K, Weber MS (2018) Functional characterization of reappearing B cells after anti-CD20 treatment of CNS autoimmune disease. Proc Natl Acad Sci U S A 115(39):9773–9778. https://doi.org/10.1073/pnas.1810470115
Huck C, Leppert D, Wegert V, Schmid C, Dunn R, Weckbecker G, Smith PA (2019) Low-dose subcutaneous anti-CD20 treatment depletes disease relevant B cell subsets and attenuates neuroinflammation. J NeuroImmune Pharmacol 14(4):709–719. https://doi.org/10.1007/s11481-019-09872-z
Curran CS, Gupta S, Sanz I, Sharon E (2019) PD-1 immunobiology in systemic lupus erythematosus. J Autoimmun 97:1–9. https://doi.org/10.1016/j.jaut.2018.10.025
Shindo Y, Yoshimura K, Kuramasu A, Watanabe Y, Ito H, Kondo T, Oga A, Ito H, Yoshino S, Hazama S, Tamada K, Yagita H, Oka M (2015) Combination immunotherapy with 4-1BB activation and PD-1 blockade enhances antitumor efficacy in a mouse model of subcutaneous tumor. Anticancer Res 35(1):129–136
Kamburova EG, Koenen HJ, Boon L, Hilbrands LB, Joosten I (2012) In vitro effects of rituximab on the proliferation, activation and differentiation of human B cells. Am J Transplant 12(2):341–350. https://doi.org/10.1111/j.1600-6143.2011.03833.x
Arbuckle MR, McClain MT, Rubertone MV, Scofield RH, Dennis GJ, James JA et al (2003) Development of autoantibodies before the clinical onset of systemic lupus erythematosus. New Engl J Med 349(16):1526–1533. https://doi.org/10.1056/NEJMoa021933
Keir ME, Butte MJ, Freeman GJ, Sharpe AH (2008) PD-1 and its ligands in tolerance and immunity. Annu Rev Immunol 26:677–704. https://doi.org/10.1146/annurev.immunol.26.021607.090331
The authors wish to thank Chuanwang Song, Hongtao Wang, Jie Tang for their direction and supervision during the experiment.
Availability of data and materials
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
Research was supported by funding from the Anhui Provincial Natural Science Foundation (1608085MH215), the Key Project of the Natural Science Foundation Universities Anhui Province (KJ2019A0319, KJ2016A475), the Natural Science Foundation of Universities in Anhui Province (KJ2013B139), the Postgraduate Science and Technology Innovation Project of Bengbu Medical College in Anhui Province (Byycx1824), and the Undergraduate Training Programs for Innovation and Entrepreneurship in Anhui Province (201,810,367,017, and 201,810,367,056).
Ethics approval and consent to participate
All participants provided informed written consent. This study was approved by the institutional review board of the First Affiliated Hospital of Bengbu Medical College.
Conflict of interest
The authors declare no financially oriented conflicts of interest. Ethical approvals and informed written consent were obtained and provided from and to all participants. The study design was approved by the Institutional Review Board of the First Affiliated Hospital of Bengbu Medical College.
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Zhu, Q., Li, Y., Zhang, L. et al. Patients with systemic lupus erythematosus show increased proportions of CD19+CD20− B cells and secretion of related autoantibodies. Clin Rheumatol (2020). https://doi.org/10.1007/s10067-020-05220-2
- B cell
- Surface marker
- Systemic lupus erythematosus