The proteasome inhibitor bortezomib depletes plasma cells and protects mice with lupus-like disease from nephritis

Abstract

Autoantibody-mediated diseases like myasthenia gravis, autoimmune hemolytic anemia and systemic lupus erythematosus represent a therapeutic challenge. In particular, long-lived plasma cells producing autoantibodies resist current therapeutic and experimental approaches. Recently, we showed that the sensitivity of myeloma cells toward proteasome inhibitors directly correlates with their immunoglobulin synthesis rates. Therefore, we hypothesized that normal plasma cells are also hypersensitive to proteasome inhibition owing to their extremely high amount of protein biosynthesis. Here we show that the proteasome inhibitor bortezomib, which is approved for the treatment of multiple myeloma, eliminates both short- and long-lived plasma cells by activation of the terminal unfolded protein response. Treatment with bortezomib depleted plasma cells producing antibodies to double-stranded DNA, eliminated autoantibody production, ameliorated glomerulonephritis and prolonged survival of two mouse strains with lupus-like disease, NZB/W F1 and MRL/lpr mice. Hence, the elimination of autoreactive plasma cells by proteasome inhibitors might represent a new treatment strategy for antibody-mediated diseases.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Elimination of plasma cells from spleens and bone marrows by bortezomib treatment in NZB/W F1 mice.
Figure 2: Cellular effects after one-week treatment with bortezomib, cylophosphamide or dexamethasone in NZB/W F1 mice.
Figure 3: Bortezomib treatment activates the terminal UPR.
Figure 4: Bortezomib treatment prevents and ameliorates lupus-like disease in NZB/W F1 mice.
Figure 5: Bortezomib (Bz) treatment ameliorates lupus-like disease in MRL/lpr mice.

References

  1. 1

    Lipsky, P.E. Systemic lupus erythematosus: an autoimmune disease of B cell hyperactivity. Nat. Immunol. 2, 764–766 (2001).

    CAS  Article  Google Scholar 

  2. 2

    Mills, J.A. Systemic lupus erythematosus. N. Engl. J. Med. 330, 1871–1879 (1994).

    CAS  Article  Google Scholar 

  3. 3

    Manz, R.A. & Radbruch, A. Plasma cells for a lifetime? Eur. J. Immunol. 32, 923–927 (2002).

    CAS  Article  Google Scholar 

  4. 4

    Miller, J.J., III & Cole, L.J. Resistance of long-lived lymphocytes and plasma cells in rat lymph nodes to treatment with prednisone, cyclophosphamide, 6 mercaptopurine and actinomycin D. J. Exp. Med. 126, 109–125 (1967).

    CAS  Article  Google Scholar 

  5. 5

    Slifka, M.K. & Ahmed, R. Long-lived plasma cells: a mechanism for maintaining persistent antibody production. Curr. Opin. Immunol. 10, 252–258 (1998).

    CAS  Article  Google Scholar 

  6. 6

    Ang, H.A., Apperley, J.F. & Ward, K.N. Persistence of antibody to human parvovirus B19 after allogeneic bone marrow transplantation: role of prior recipient immunity. Blood 89, 4646–4651 (1997).

    CAS  PubMed  Google Scholar 

  7. 7

    van Tol, M.J. et al. The origin of IgG production and homogeneous IgG components after allogeneic bone marrow transplantation. Blood 87, 818–826 (1996).

    CAS  PubMed  Google Scholar 

  8. 8

    Theofilopoulos, A.N. & Dixon, F.J. Murine models of systemic lupus erythematosus. Adv. Immunol. 37, 269–390 (1985).

    CAS  Article  Google Scholar 

  9. 9

    Hoyer, B.F. et al. Short-lived plasmablasts and long-lived plasma cells contribute to chronic humoral autoimmunity in NZB/W mice. J. Exp. Med. 199, 1577–1584 (2004).

    CAS  Article  Google Scholar 

  10. 10

    Manz, R.A., Hauser, A.E., Hiepe, F. & Radbruch, A. Maintenance of serum antibody levels. Annu. Rev. Immunol. 23, 367–386 (2005).

    CAS  Article  Google Scholar 

  11. 11

    Adams, J. The proteasome: a suitable antineoplastic target. Nat. Rev. Cancer 4, 349–360 (2004).

    CAS  Article  Google Scholar 

  12. 12

    Hideshima, T., Richardson, P.G. & Anderson, K.C. Targeting proteasome inhibition in hematologic malignancies. Rev. Clin. Exp. Hematol. 7, 191–204 (2003).

    CAS  PubMed  Google Scholar 

  13. 13

    Rajkumar, S.V., Richardson, P.G., Hideshima, T. & Anderson, K.C. Proteasome inhibition as a novel therapeutic target in human cancer. J. Clin. Oncol. 23, 630–639 (2005).

    CAS  Article  Google Scholar 

  14. 14

    Meister, S. et al. Extensive immunoglobulin production sensitizes myeloma cells for proteasome inhibition. Cancer Res. 67, 1783–1792 (2007).

    CAS  Article  Google Scholar 

  15. 15

    Obeng, E.A. et al. Proteasome inhibitors induce a terminal unfolded protein response in multiple myeloma cells. Blood 107, 4907–4916 (2006).

    CAS  Article  Google Scholar 

  16. 16

    Hibi, T. & Dosch, H.M. Limiting dilution analysis of the B cell compartment in human bone marrow. Eur. J. Immunol. 16, 139–145 (1986).

    CAS  Article  Google Scholar 

  17. 17

    Lefkowith, J.B. & Gilkeson, G.S. Nephritogenic autoantibodies in lupus: current concepts and continuing controversies. Arthritis Rheum. 39, 894–903 (1996).

    CAS  Article  Google Scholar 

  18. 18

    Hahn, B.H. Antibodies to DNA. N. Engl. J. Med. 338, 1359–1368 (1998).

    CAS  Article  Google Scholar 

  19. 19

    Manz, R.A., Thiel, A. & Radbruch, A. Lifetime of plasma cells in the bone marrow. Nature 388, 133–134 (1997).

    CAS  Article  Google Scholar 

  20. 20

    Radbruch, A. et al. Competence and competition: the challenge pf becoming a long-lived plasma cell. Nat. Rev. Immunol. 6, 741–750 (2006).

    CAS  Article  Google Scholar 

  21. 21

    Nencioni, A. et al. Proteasome inhibitor bortezomib modulates TLR4-induced dendritic cell activation. Blood 108, 551–558 (2006).

    CAS  Article  Google Scholar 

  22. 22

    Straube, C. et al. Bortezomib significantly impairs the immunostimulatory capacity of human myeloid blood dendritic cells. Leukemia 21, 1464–1471 (2007).

    CAS  Article  Google Scholar 

  23. 23

    Oyadomari, S. & Mori, M. Roles of CHOP/GADD153 in endoplasmic reticulum stress. Cell Death Differ. 11, 381–389 (2004).

    CAS  Article  Google Scholar 

  24. 24

    Moran, S.T. et al. Synergism between NF-κB1/p50 and Notch2 during the development of marginal zone B lymphocytes. J. Immunol. 179, 195–200 (2007).

    CAS  Article  Google Scholar 

  25. 25

    Sasaki, Y. et al. Canonical NF-κB activity, dispensable for B cell development, replaces BAFF-receptor signals and promotes B cell proliferation upon activation. Immunity 24, 729–739 (2006).

    CAS  Article  Google Scholar 

  26. 26

    Maseda, D., Meister, S., Neubert, K., Herrmann, M. & Voll, R.E. Proteasome inhibition drastically but reversibly impairs murine lymphocyte development. Cell Death Differ. 15, 600–612 (2008).

    CAS  Article  Google Scholar 

  27. 27

    Schubert, U. et al. Rapid degradation of a large fraction of newly synthesized proteins by proteasomes. Nature 404, 770–774 (2000).

    CAS  Article  Google Scholar 

  28. 28

    Gass, J.N., Gifford, N.M. & Brewer, J.W. Activation of an unfolded protein response during differentiation of antibody-secreting B cells. J. Biol. Chem. 277, 49047–49054 (2002).

    CAS  Article  Google Scholar 

  29. 29

    Iwakoshi, N.N. et al. Plasma cell differentiation and the unfolded protein response intersect at the transcription factor XBP-1. Nat. Immunol. 4, 321–329 (2003).

    CAS  Article  Google Scholar 

  30. 30

    Kaufman, R.J. Stress signaling from the lumen of the endoplasmic reticulum: coordination of gene transcriptional and translational controls. Genes Dev. 13, 1211–1233 (1999).

    CAS  Article  Google Scholar 

  31. 31

    Patil, C. & Walter, P. Intracellular signaling from the endoplasmic reticulum to the nucleus: the unfolded protein response in yeast and mammals. Curr. Opin. Cell Biol. 13, 349–355 (2001).

    CAS  Article  Google Scholar 

  32. 32

    Reimold, A.M. et al. Plasma cell differentiation requires the transcription factor XBP-1. Nature 412, 300–307 (2001).

    CAS  Article  Google Scholar 

  33. 33

    van Anken, E. et al. Sequential waves of functionally related proteins are expressed when B cells prepare for antibody secretion. Immunity 18, 243–253 (2003).

    CAS  Article  Google Scholar 

  34. 34

    Wiertz, E.J. et al. Sec61-mediated transfer of a membrane protein from the endoplasmic reticulum to the proteasome for destruction. Nature 384, 432–438 (1996).

    CAS  Article  Google Scholar 

  35. 35

    Kim, R., Emi, M., Tanabe, K. & Murakami, S. Role of the unfolded protein response in cell death. Apoptosis 11, 5–13 (2006).

    CAS  Article  Google Scholar 

  36. 36

    Nawrocki, S.T. et al. Bortezomib inhibits PKR-like endoplasmic reticulum (ER) kinase and induces apoptosis via ER stress in human pancreatic cancer cells. Cancer Res. 65, 11510–11519 (2005).

    CAS  Article  Google Scholar 

  37. 37

    Nawrocki, S.T. et al. Bortezomib sensitizes pancreatic cancer cells to endoplasmic reticulum stress-mediated apoptosis. Cancer Res. 65, 11658–11666 (2005).

    CAS  Article  Google Scholar 

  38. 38

    Schaumann, D.H., Tuischer, J., Ebell, W., Manz, R.A. & Lauster, R. VCAM-1–positive stromal cells from human bone marrow producing cytokines for B lineage progenitors and for plasma cells: SDF-1, flt3L and BAFF. Mol. Immunol. 44, 1606–1612 (2007).

    CAS  Article  Google Scholar 

  39. 39

    Tokoyoda, K., Egawa, T., Sugiyama, T., Choi, B.I. & Nagasawa, T. Cellular niches controlling B lymphocyte behavior within bone marrow during development. Immunity 20, 707–718 (2004).

    CAS  Article  Google Scholar 

  40. 40

    Klinman, D.M. & Steinberg, A.D. Inquiry into murine and human lupus. Immunol. Rev. 144, 157–193 (1995).

    CAS  Article  Google Scholar 

  41. 41

    Winfield, J.B., Faiferman, I. & Koffler, D. Avidity of anti-DNA antibodies in serum and IgG glomerular eluates from patients with systemic lupus erythematosus. Association of high avidity antinative DNA antibody with glomerulonephritis. J. Clin. Invest. 59, 90–96 (1977).

    CAS  Article  Google Scholar 

  42. 42

    Cambridge, G. et al. B cell depletion therapy in systemic lupus erythematosus: effect on autoantibody and antimicrobial antibody profiles. Arthritis Rheum. 54, 3612–3622 (2006).

    CAS  Article  Google Scholar 

  43. 43

    Sfikakis, P.P. et al. Remission of proliferative lupus nephritis following B cell depletion therapy is preceded by down-regulation of the T cell costimulatory molecule CD40 ligand: an open-label trial. Arthritis Rheum. 52, 501–513 (2005).

    CAS  Article  Google Scholar 

  44. 44

    Leandro, M.J., Edwards, J.C., Cambridge, G., Ehrenstein, M.R. & Isenberg, D.A. An open study of B lymphocyte depletion in systemic lupus erythematosus. Arthritis Rheum. 46, 2673–2677 (2002).

    Article  Google Scholar 

  45. 45

    Emery, P. et al. The efficacy and safety of rituximab in patients with active rheumatoid arthritis despite methotrexate treatment: results of a phase IIB randomized, double-blind, placebo-controlled, dose-ranging trial. Arthritis Rheum. 54, 1390–1400 (2006).

    CAS  Article  Google Scholar 

  46. 46

    Looney, R.J. et al. B cell depletion as a novel treatment for systemic lupus erythematosus: a phase I/II dose-escalation trial of rituximab. Arthritis Rheum. 50, 2580–2589 (2004).

    CAS  Article  Google Scholar 

  47. 47

    Medina, F., Segundo, C., Campos-Caro, A., Gonzalez-Garcia, I. & Brieva, J.A. The heterogeneity shown by human plasma cells from tonsil, blood, and bone marrow reveals graded stages of increasing maturity, but local profiles of adhesion molecule expression. Blood 99, 2154–2161 (2002).

    CAS  Article  Google Scholar 

  48. 48

    Benner, R., Van Oudenaren, A. & Koch, G. Induction of antibody formation in mouse bone marrow. in Immunological Methods Vol. II (eds. Pernis, B. & Lefkovits, I.) 247–262 (Academic Press, New York, 1981).

    Google Scholar 

  49. 49

    Wellmann, U., Letz, M., Schneider, A., Amann, K. & Winkler, T.H. An Ig μ-heavy chain transgene inhibits systemic lupus erythematosus immunopathology in autoimmune (NZB x NZW)F1 mice. Int. Immunol. 13, 1461–1469 (2001).

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We are grateful to U. Appelt for expert cell sorting and M. Wiesener and F. Nimmerjahn for critical reading the manuscript. This work was supported by the Interdisciplinary Center for Clinical Research (project number N2) and the German Research Society (project VO673/31 and Collaborative Research Centers SFB 643; project B3, both to R.E.V.). Parts of this work were funded by an intramural grant from the ELAN fond, a Training Grant GK 592 from the German Research Society and Collaborative Research Centers SFB 423 (project Z2).

Author information

Affiliations

Authors

Contributions

K.N. designed experiments; performed animal studies, flow cytometry, ELISAs, ELISPOTs and quantitative real-time RT-PCR; analyzed data; generated figures and wrote the manuscript. S.M. assisted in performing flow cytometry and animal experiments and participated in discussions. K.M. performed and assisted with the BrdU experiments, provided antibodies and participated in discussions. F.W. performed RNA isolation and cDNA synthesis and participated in discussions. D.M. assisted with the animal studies. K.A. performed all histological analyses. C.W. contributed to the analyses of DC and T cell function. T.H.W. provided crucial ideas, participated in discussions and edited the manuscript. J.R.K. participated in discussions and edited the manuscript. R.A.M. crucially participated in design and analyses of the BrdU labeling experiments, provided antibodies, participated in discussions and edited the manuscript. R.E.V. provided crucial ideas, designed the study and experiments, supervised the study and wrote the manuscript together with K.N.

Corresponding author

Correspondence to Reinhard E Voll.

Ethics declarations

Competing interests

R.E.V. holds stocks of Millenium Pharmaceuticals, the manufacturer of bortezomib (value of less than $15,000).

K.N., S.M., K.M., R.A.M., D.M., J.R.K. and R.E.V. are inventors listed on a patent application for the use of proteasome inhibitors to deplete plasma cells.

Supplementary information

Supplementary Text and Figures

Supplementary Figs. 1–9 and Supplementary Methods (PDF 227 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Neubert, K., Meister, S., Moser, K. et al. The proteasome inhibitor bortezomib depletes plasma cells and protects mice with lupus-like disease from nephritis. Nat Med 14, 748–755 (2008). https://doi.org/10.1038/nm1763

Download citation

Further reading

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing