Rheumatoid Arthritis
Modulation of molecular imprints in the antigen-experienced B cell repertoire by rituximab
Arumugam Palanichamy, Petra Roll, Regina Theiss, Thomas Dörner,
Hans-Peter Tony,
Thomas Dörner
Charité Universitätsmedizin Berlin, University of Berlin, Berlin, Germany
Dr. Dörner has received consulting fees and speaking fees from Roche and Genentech (less than $10,000 each).
Search for more papers by this authorCorresponding Author
Hans-Peter Tony
University of Würzburg, Würzburg, Germany
Dr. Tony has received speaking fees from Roche, Abbott, Essex, and Wyeth (less than $10,000 each).
Medizinische Klinik II, Department of Rheumatology/Clinical Immunology, Klinikstrasse 6, 97070 Würzburg, GermanySearch for more papers by this authorArumugam Palanichamy, Petra Roll, Regina Theiss, Thomas Dörner,
Hans-Peter Tony,
Thomas Dörner
Charité Universitätsmedizin Berlin, University of Berlin, Berlin, Germany
Dr. Dörner has received consulting fees and speaking fees from Roche and Genentech (less than $10,000 each).
Search for more papers by this authorCorresponding Author
Hans-Peter Tony
University of Würzburg, Würzburg, Germany
Dr. Tony has received speaking fees from Roche, Abbott, Essex, and Wyeth (less than $10,000 each).
Medizinische Klinik II, Department of Rheumatology/Clinical Immunology, Klinikstrasse 6, 97070 Würzburg, GermanySearch for more papers by this authorAbstract
Objective
Transient B cell depletion by rituximab has recently gained more importance in the treatment of rheumatic disorders. Nevertheless, little is known about the reemerging B cells. We analyzed dynamic changes in the repopulating B cells, particularly the postswitch B cells, and studied the mutational patterns of Ig genes in antigen-experienced B cells.
Methods
Five patients with active rheumatoid arthritis (RA) were treated with rituximab. In 3 patients, B cell receptor (BCR) gene analysis was performed before treatment and during B cell recovery using genomic DNA. In 2 patients, B cell subsets were studied during the early recovery phase using single-cell technology. For comparison, immunophenotyping of B cell subsets was performed.
Results
Early B cell recovery was marked by a relatively expanded population of highly mutated B cells, which were correlated with B cells with a plasmablast phenotype on comparative immunophenotyping. Analysis of the mutational pattern in these cells revealed increased RGYW/WRCY (where R = A/G, Y = C/T, and W = A/T) hotspot targeting (44% before rituximab versus 59% after) and elevated ratios of replacement to silent mutations within the complementarity-determining regions in Ig genes (1.87 before rituximab versus 2.67 after; P ≤ 0.0025).
Conclusion
Our findings show that rituximab leads to qualitative changes in the imprints of highly mutated, antigen-experienced BCRs, representing the result of selection, whereas molecular processes such as Ig V rearrangements are not affected by this treatment.
REFERENCES
- 1 Lipsky PE. Systemic lupus erythematosus: an autoimmune disease of B cell hyperactivity. Nat Immunol 2001; 2: 764–6.
- 2 Mariette X. The B cell: a new therapeutic target in rheumatoid arthritis and other autoimmune diseases. Joint Bone Spine 2004; 71: 357–60.
- 3 Silverman GJ, Weisman S. Rituximab therapy and autoimmune disorders: prospects for anti-B cell therapy [review]. Arthritis Rheum 2003; 48: 1484–92.
- 4 Kuwana M, Nomura S, Fujimura K, Nagasawa T, Muto Y, Kurata Y, et al. Effect of a single injection of humanized anti-CD154 monoclonal antibody on the platelet-specific autoimmune response in patients with immune thrombocytopenic purpura. Blood 2004; 103: 1229–36.
- 5 Halpern WG, Lappin P, Zanardi T, Cai W, Corcoran M, Zhong Z, et al. Chronic administration of belimumab, a BLyS antagonist, decreases tissue and peripheral blood B-lymphocyte populations in cynomolgus monkeys: pharmacokinetic, pharmacodynamic, and toxicologic effects. Toxicol Sci 2006; 91: 586–99.
- 6 Kalunian KC, Davis JC Jr, Merrill JT, Totoritis MC, Wofsy D, for the IDEC-131 Lupus Study Group. Treatment of systemic lupus erythematosus by inhibition of T cell costimulation with anti-CD154: a randomized, double-blind, placebo-controlled trial. Arthritis Rheum 2002; 46: 3251–8.
- 7 Vugmeyster Y, Seshasayee D, Chang W, Storn A, Howell K, Sa S, et al. A soluble BAFF antagonist, BR3-Fc, decreases peripheral blood B cells and lymphoid tissue marginal zone and follicular B cells in cynomolgus monkeys. Am J Pathol 2006; 168: 476–89.
- 8 Browning JL. B cells move to centre stage: novel opportunities for autoimmune disease treatment. Nat Rev Drug Discov 2006; 5: 564–76.
- 9 Kneitz C, Wilhelm M, Tony HP. Effective B cell depletion with rituximab in the treatment of autoimmune diseases. Immunobiology 2002; 206: 519–27.
- 10 Keystone E. B cell targeted therapies. Arthritis Res Ther 2005; 7 Suppl 3: S13–8.
- 11 Eisenberg R. Do autoantigens define autoimmunity or vice versa? Eur J Immunol 2005; 35: 367–70.
- 12 Hamaguchi Y, Uchida J, Cain DW, Venturi GM, Poe JC, Haas KM, et al. The peritoneal cavity provides a protective niche for B1 and conventional B lymphocytes during anti-CD20 immunotherapy in mice. J Immunol 2005; 174: 4389–99.
- 13 Gong Q, Ou Q, Ye S, Lee WP, Cornelius J, Diehl L, et al. Importance of cellular microenvironment and circulatory dynamics in B cell immunotherapy. J Immunol 2005; 174: 817–26.
- 14 Looney RJ, Anolik J, Sanz I. B lymphocytes in systemic lupus erythematosus: lessons from therapy targeting B cells. Lupus 2004; 13: 381–90.
- 15 Leandro MJ, Cooper N, Cambridge G, Ehrenstein MR, Edwards JC. Bone marrow B-lineage cells in patients with rheumatoid arthritis following rituximab therapy. Rheumatology (Oxford) 2007; 46: 29–36.
- 16 Leandro MJ, Cambridge G, Ehrenstein MR, Edwards JC. Reconstitution of peripheral blood B cells after depletion with rituximab in patients with rheumatoid arthritis. Arthritis Rheum 2006; 54: 613–20.
- 17 Roll P, Palanichamy A, Kneitz C, Dorner T, Tony HP. Regeneration of B cell subsets after transient B cell depletion using anti-CD20 antibodies in rheumatoid arthritis. Arthritis Rheum 2006; 54: 2377–86.
- 18 Sanz I, Anolik J. Reconstitution of the adult B cell repertoire after treatment with rituximab. Arthritis Res Ther 2005; 7: 175–6.
- 19 Anolik JH, Friedberg JW, Zheng B, Barnard J, Owen T, Cushing E, et al. B cell reconstitution after rituximab treatment of lymphoma recapitulates B cell ontogeny. Clin Immunol 2007; 122: 139–45.
- 20 Odegard VH, Schatz DG. Targeting of somatic hypermutation. Nat Rev Immunol 2006; 6: 573–83.
- 21 Balazs M, Martin F, Zhou T, Kearney J. Blood dendritic cells interact with splenic marginal zone B cells to initiate T-independent immune responses. Immunity 2002; 17: 341–52.
- 22 William J, Euler C, Christensen S, Shlomchik MJ. Evolution of autoantibody responses via somatic hypermutation outside of germinal centers. Science 2002; 297: 2066–70.
- 23 Tiller T, Tsuiji M, Yurasov S, Velinzon K, Nussenzweig MC, Wardemann H. Autoreactivity in human IgG+ memory B cells. Immunity 2007; 26: 205–13.
- 24 Arnett FC, Edworthy SM, Bloch DA, McShane DJ, Fries JF, Cooper NS, et al. The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum 1988; 31: 315–24.
- 25 Prevoo ML, van ‘t Hof MA, Kuper HH, van Leeuwen MA, van de Putte LB, van Riel PL. Modified disease activity scores that include twenty-eight–joint counts: development and validation in a prospective longitudinal study of patients with rheumatoid arthritis. Arthritis Rheum 1995; 38: 44–8.
- 26 Brezinschek HP, Brezinschek RI, Lipsky PE. Analysis of the heavy chain repertoire of human peripheral B cells using single-cell polymerase chain reaction. J Immunol 1995; 155: 190–202.
- 27 Dorner T, Brezinschek HP, Brezinschek RI, Foster SJ, Domiati-Saad R, Lipsky PE. Analysis of the frequency and pattern of somatic mutations within nonproductively rearranged human variable heavy chain genes. J Immunol 1997; 158: 2779–89.
- 28 Souto-Carneiro MM, Longo NS, Russ DE, Sun HW, Lipsky PE. Characterization of the human Ig heavy chain antigen binding complementarity determining region 3 using a newly developed software algorithm, JOINSOLVER. J Immunol 2004; 172: 6790–802.
- 29 Ruzickova S, Pruss A, Odendahl M, Wolbart K, Burmester GR, Scholze J, et al. Chronic lymphocytic leukemia preceded by cold agglutinin disease: intraclonal immunoglobulin light-chain diversity in VH4-34 expressing single leukemic B cells [published erratum appears in Blood 2003;101:1676]. Blood 2002; 100: 3419–22.
- 30
Dorner T,
Lipsky PE.
Immunoglobulin variable-region gene usage in systemic autoimmune diseases [review].
Arthritis Rheum
2001;
44:
2715–27.
10.1002/1529-0131(200112)44:12<2715::AID-ART458>3.0.CO;2-L CAS PubMed Web of Science® Google Scholar
- 31 Dorner T, Foster SJ, Brezinschek HP, Lipsky PE. Analysis of the targeting of the hypermutational machinery and the impact of subsequent selection on the distribution of nucleotide changes in human VHDJH rearrangements. Immunol Rev 1998; 162: 161–71.
- 32 Farner NL, Dorner T, Lipsky PE. Molecular mechanisms and selection influence the generation of the human VλJλ repertoire. J Immunol 1999; 162: 2137–45.
- 33 Anolik JH, Barnard J, Cappione A, Pugh-Bernard AE, Felgar RE, Looney RJ, et al. Rituximab improves peripheral B cell abnormalities in human systemic lupus erythematosus. Arthritis Rheum 2004; 50: 3580–90.
- 34 Silverman GJ. Therapeutic B cell depletion and regeneration in rheumatoid arthritis: emerging patterns and paradigms [review]. Arthritis Rheum 2006; 54: 2356–67.
- 35 Cohen SB, Emery P, Greenwald MW, Dougados M, Furie RA, Genovese MC, et al, for the REFLEX Trial Group. Rituximab for rheumatoid arthritis refractory to anti–tumor necrosis factor therapy: results of a multicenter, randomized, double-blind, placebo-controlled, phase III trial evaluating primary efficacy and safety at twenty-four weeks. Arthritis Rheum 2006; 54: 2793–806.
- 36 Leandro MJ, Cambridge G, Edwards JC, Ehrenstein MR, Isenberg DA. B-cell depletion in the treatment of patients with systemic lupus erythematosus: a longitudinal analysis of 24 patients. Rheumatology (Oxford) 2005; 44: 1542–5.
- 37 Storek J, Ferrara S, Ku N, Giorgi JV, Champlin RE, Saxon A. B cell reconstitution after human bone marrow transplantation: recapitulation of ontogeny? Bone Marrow Transplant 1993; 12: 387–98.
- 38 Avanzini MA, Locatelli F, Dos Santos C, Maccario R, Lenta E, Oliveri M, et al. B lymphocyte reconstitution after hematopoietic stem cell transplantation: functional immaturity and slow recovery of memory CD27+ B cells. Exp Hematol 2005; 33: 480–6.
- 39 Rouziere AS, Kneitz C, Palanichamy A, Dorner T, Tony HP. Regeneration of the immunoglobulin heavy-chain repertoire after transient B-cell depletion with an anti-CD20 antibody. Arthritis Res Ther 2005; 7: R714–24.
- 40 Kim HJ, Berek C. B cells in rheumatoid arthritis. Arthritis Res 2000; 2: 126–31.
- 41
Cassese G,
Lindenau S,
de Boer B,
Arce S,
Hauser A,
Riemekasten G, et al.
Inflamed kidneys of NZB/W mice are a major site for the homeostasis of plasma cells.
Eur J Immunol
2001;
31:
2726–32.
10.1002/1521-4141(200109)31:9<2726::AID-IMMU2726>3.0.CO;2-H CAS PubMed Web of Science® Google Scholar
- 42 Hoyer BF, Moser K, Hauser AE, Peddinghaus A, Voigt C, Eilat D, et al. Short-lived plasmablasts and long-lived plasma cells contribute to chronic humoral autoimmunity in NZB/W mice. J Exp Med 2004; 199: 1577–84.
- 43 Brezinschek HP, Dorner T, Monson NL, Brezinschek RI, Lipsky PE. The influence of CD40-CD154 interactions on the expressed human VH repertoire: analysis of VH genes expressed by individual B cells of a patient with X-linked hyper-IgM syndrome. Int Immunol 2000; 12: 767–75.
- 44
Foster SJ,
Dorner T,
Lipsky PE.
Targeting and subsequent selection of somatic hypermutations in the human Vκ repertoire.
Eur J Immunol
1999;
29:
3122–32.
10.1002/(SICI)1521-4141(199910)29:10<3122::AID-IMMU3122>3.0.CO;2-O CAS PubMed Web of Science® Google Scholar
- 45
Monson NL,
Dorner T,
Lipsky PE.
Targeting and selection of mutations in human Vλ rearrangements.
Eur J Immunol
2000;
30:
1597–605.
10.1002/1521-4141(200006)30:6<1597::AID-IMMU1597>3.0.CO;2-R CAS PubMed Web of Science® Google Scholar
- 46 Randen I, Mellbye OJ, Forre O, Natvig JB. The identification of germinal centres and follicular dendritic cell networks in rheumatoid synovial tissue. Scand J Immunol 1995; 41: 481–6.
- 47 Stott DI, Hiepe F, Hummel M, Steinhauser G, Berek C. Antigen-driven clonal proliferation of B cells within the target tissue of an autoimmune disease: the salivary glands of patients with Sjögren’s syndrome. J Clin Invest 1998; 102: 938–46.
