Article

Plasma cells require autophagy for sustainable immunoglobulin production

  • Nature Immunology volume 14, pages 298305 (2013)
  • doi:10.1038/ni.2524
  • Download Citation
Received:
Accepted:
Published:

Abstract

The role of autophagy in plasma cells is unknown. Here we found notable autophagic activity in both differentiating and long-lived plasma cells and investigated its function through the use of mice with conditional deficiency in the essential autophagic molecule Atg5 in B cells. Atg5−/− differentiating plasma cells had a larger endoplasmic reticulum (ER) and more ER stress signaling than did their wild-type counterparts, which led to higher expression of the transcriptional repressor Blimp-1 and immunoglobulins and more antibody secretion. The enhanced immunoglobulin synthesis was associated with less intracellular ATP and more death of mutant plasma cells, which identified an unsuspected autophagy-dependent cytoprotective trade-off between immunoglobulin synthesis and viability. In vivo, mice with conditional deficiency in Atg5 in B cells had defective antibody responses, complete selection in the bone marrow for plasma cells that escaped Atg5 deletion and fewer antigen-specific long-lived bone marrow plasma cells than did wild-type mice, despite having normal germinal center responses. Thus, autophagy is specifically required for plasma cell homeostasis and long-lived humoral immunity.

  • Subscribe to Nature Immunology for full access:

    $59

    Subscribe

Additional access options:

Already a subscriber?  Log in  now or  Register  for online access.

References

  1. 1.

    , , , & The genetic network controlling plasma cell differentiation. Semin. Immunol. 23, 341–349 (2011).

  2. 2.

    & Signal integration in the endoplasmic reticulum unfolded protein response. Nat. Rev. Mol. Cell Biol. 8, 519–529 (2007).

  3. 3.

    & Regulation of plasma-cell development. Nat. Rev. Immunol. 5, 230–242 (2005).

  4. 4.

    , & On the redox control of B lymphocyte differentiation and function. Antioxid. Redox Signal. 16, 1139–1149 (2012).

  5. 5.

    et al. Progressively impaired proteasomal capacity during terminal plasma cell differentiation. EMBO J. 25, 1104–1113 (2006).

  6. 6.

    et al. Dampening Ab responses using proteasome inhibitors following in vivo B cell activation. Eur. J. Immunol. 38, 658–667 (2008).

  7. 7.

    et al. The proteasome inhibitor bortezomib depletes plasma cells and protects mice with lupus-like disease from nephritis. Nat. Med. 14, 748–755 (2008).

  8. 8.

    , & Autophagy in metazoans: cell survival in the land of plenty. Nat. Rev. Mol. Cell Biol. 6, 439–448 (2005).

  9. 9.

    & Regulation mechanisms and signaling pathways of autophagy. Annu. Rev. Genet. 43, 67–93 (2009).

  10. 10.

    , & Autophagy and the integrated stress response. Mol. Cell 40, 280–293 (2010).

  11. 11.

    , & Autophagy counterbalances endoplasmic reticulum expansion during the unfolded protein response. PLoS Biol. 4, e423 (2006).

  12. 12.

    et al. Two endoplasmic reticulum-associated degradation (ERAD) systems for the novel variant of the mutant dysferlin: ubiquitin/proteasome ERAD(I) and autophagy/lysosome ERAD(II). Hum. Mol. Genet. 16, 618–629 (2007).

  13. 13.

    et al. Autophagic elimination of misfolded procollagen aggregates in the endoplasmic reticulum as a means of cell protection. Mol. Biol. Cell 20, 2744–2754 (2009).

  14. 14.

    Autophagy in immunity and cell-autonomous defense against intracellular microbes. Immunol. Rev. 240, 92–104 (2011).

  15. 15.

    , & Autophagy in immunity and inflammation. Nature 469, 323–335 (2011).

  16. 16.

    et al. The autophagy gene ATG5 plays an essential role in B lymphocyte development. Autophagy 4, 309–314 (2008).

  17. 17.

    et al. Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy 8, 445–544 (2012).

  18. 18.

    , , , & In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker. Mol. Biol. Cell 15, 1101–1111 (2004).

  19. 19.

    & Selective autophagy mediated by autophagic adapter proteins. Autophagy 7, 279–296 (2011).

  20. 20.

    , , , & Dynamic and reversible restructuring of the ER induced by PDMP in cultured cells. J. Cell Sci. 119, 3249–3260 (2006).

  21. 21.

    , , & The unfolded protein response of B-lymphocytes: PERK-independent development of antibody-secreting cells. Mol. Immunol. 45, 1035–1043 (2008).

  22. 22.

    , , , & Plasma cell differentiation initiates a limited ER stress response by specifically suppressing the PERK-dependent branch of the unfolded protein response. Cell Stress Chaperones 15, 281–293 (2010).

  23. 23.

    et al. Blimp-1 orchestrates plasma cell differentiation by extinguishing the mature B cell gene expression program. Immunity 17, 51–62 (2002).

  24. 24.

    , & BLIMP-1 is a target of cellular stress and downstream of the unfolded protein response. Eur. J. Immunol. 36, 1572–1582 (2006).

  25. 25.

    et al. Metabolomics of B to plasma cell differentiation. J. Proteome Res. 10, 4165–4176 (2011).

  26. 26.

    & Vagaries of conditional gene targeting. Nat. Immunol. 8, 665–668 (2007).

  27. 27.

    , , & B cell clones that sustain long-term plasmablast growth in T-independent extrafollicular antibody responses. Proc. Natl. Acad. Sci. USA 103, 5905–5910 (2006).

  28. 28.

    , & Antigen-loading compartments for major histocompatibility complex class II molecules continuously receive input from autophagosomes. Immunity 26, 79–92 (2007).

  29. 29.

    et al. In vivo requirement for Atg5 in antigen presentation by dendritic cells. Immunity 32, 227–239 (2010).

  30. 30.

    , & Proteostenosis and plasma cell pathophysiology. Curr. Opin. Cell Biol. 23, 216–222 (2011).

  31. 31.

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

  32. 32.

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

  33. 33.

    et al. CHOP-independent apoptosis and pathway-selective induction of the UPR in developing plasma cells. Mol. Immunol. 47, 1356–1365 (2010).

  34. 34.

    , , & Protein synthesis in plasma cells is regulated by crosstalk between endoplasmic reticulum stress and mTOR signaling. Eur. J. Immunol. 41, 491–502 (2011).

  35. 35.

    Hide and Go Seek: Activation of the Secretory-Specific Poly (A) Site of Igh by Transcription Elongation Factors in RNA Processing. (ed. Grabowski, P.) 27–35 (Intech, 2011).

  36. 36.

    , & The half-life of immunoglobulin mRNA increases during B-cell differentiation: a possible role for targeting to membrane-bound polysomes. Genes Dev. 2, 1003–1011 (1988).

  37. 37.

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

  38. 38.

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

  39. 39.

    & Aiolos is required for the generation of high affinity bone marrow plasma cells responsible for long-term immunity. J. Exp. Med. 199, 209–219 (2004).

  40. 40.

    , , , & Blimp-1 is required for maintenance of long-lived plasma cells in the bone marrow. J. Exp. Med. 202, 1471–1476 (2005).

  41. 41.

    , , , & XBP-1 regulates signal transduction, transcription factors and bone marrow colonization in B cells. EMBO J. 28, 1624–1636 (2009).

  42. 42.

    et al. Improved structure, function and compatibility for CellProfiler: modular high-throughput image analysis software. Bioinformatics 27, 1179–1180 (2011).

  43. 43.

    , & Image Processing with ImageJ. Biophotonics Intern. 11, 36–42 (2004).

  44. 44.

    et al. The CXXCXXC motif determines the folding, structure and stability of human Ero1-Lα. EMBO J. 19, 4493–502 (2000).

  45. 45.

    et al. The role of bFGF on the ability of MSC to activate endogenous regenerative mechanisms in an ectopic bone formation model. Biomaterials 33, 2086–2096 (2012).

  46. 46.

    et al. Autophagy-deficient mice develop multiple liver tumors. Genes Dev. 25, 795–800 (2011).

Download references

Acknowledgements

We thank I. Braakman (University of Utrecht) for anti-PDI antiserum; N. Mizushima (Tokyo University) for GFP-LC3 mice; H.W. Virgin (Washington University) for Atg5f/fCD19-Cre mice; H. Auner, A. Bachi, M. Bertolotti, P. Cascio, P. Dellabona, F. Fontana, J. Garcia, M. Glickman, M. Iannacone, L. Maiuri, A. Manfredi, G. Merlini, A. Mondino, A. Orsi, L. Rampoldi, I. Rowe, S. Tooze, E. Tonti and E. van Anken for discussions and suggestions; T. Pengo, C. Covino and the staff of the Advanced Light and Electron Microscopy Bioimaging Center for support with microscopy; E. Canonico and I. Muradore for cell sorting; L. Spagnuolo for immunohistochemistry; and F. Loro for secretarial assistance. Supported by the Italian Ministry of Health (S. Casola; and Giovani Ricercatori 1143560 to S. Cenci), the Multiple Myeloma Research Foundation (S. Cenci), the Italian Association for Cancer Research (S. Casola; and Special Program Molecular Clinical Oncology 5 per mille 9965 to R.S. and S. Cenci), the Giovanni Armenise-Harvard Foundation Career Development Program (S. Casola) and the Italian Foundation for Cancer Research (S. Casola).

Author information

Affiliations

  1. Division of Genetics and Cell Biology, San Raffaele Scientific Institute, Milano, Italy.

    • Niccolò Pengo
    • , Maria Scolari
    • , Laura Oliva
    • , Enrico Milan
    • , Claudio Fagioli
    • , Arianna Merlini
    • , Elisabetta Mariani
    • , Elena Pasqualetto
    • , Ugo Orfanelli
    • , Roberto Sitia
    •  & Simone Cenci
  2. Università Vita-Salute San Raffaele, Milano, Italy.

    • Niccolò Pengo
    • , Enrico Milan
    • , Arianna Merlini
    • , Roberto Sitia
    •  & Simone Cenci
  3. The Institute of Molecular Oncology (IFOM) of the Italian Foundation for Cancer Research (FIRC), Milano, Italy.

    • Federica Mainoldi
    •  & Stefano Casola
  4. Imaging Research Center, San Raffaele Scientific Institute, Milano, Italy.

    • Andrea Raimondi
  5. Bone Pathophysiology Program (BoNetwork), Division of Genetics and Cell Biology, San Raffaele Scientific Institute, Milano, Italy.

    • Elisabetta Mariani
    • , Roberto Sitia
    •  & Simone Cenci
  6. Pathology Unit, San Raffaele Scientific Institute, Milano, Italy.

    • Maurilio Ponzoni
  7. Myeloma Unit, San Raffaele Scientific Institute, Milano, Italy.

    • Maurilio Ponzoni

Authors

  1. Search for Niccolò Pengo in:

  2. Search for Maria Scolari in:

  3. Search for Laura Oliva in:

  4. Search for Enrico Milan in:

  5. Search for Federica Mainoldi in:

  6. Search for Andrea Raimondi in:

  7. Search for Claudio Fagioli in:

  8. Search for Arianna Merlini in:

  9. Search for Elisabetta Mariani in:

  10. Search for Elena Pasqualetto in:

  11. Search for Ugo Orfanelli in:

  12. Search for Maurilio Ponzoni in:

  13. Search for Roberto Sitia in:

  14. Search for Stefano Casola in:

  15. Search for Simone Cenci in:

Contributions

N.P., S. Casola, R.S. and S. Cenci designed experiments; N.P., M.S., L.O., E. Milan, E.Mariani, F.M., A.R., C.F., A.M., E.P., U.O. and M.P. did the experiments; and N.P., S. Casola and S. Cenci analyzed data and wrote the paper.

Competing interests

N.P., M.S. and S.Ce. have filed a patent related to the research presented here.

Corresponding author

Correspondence to Simone Cenci.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–7 and Table 1

Excel files

  1. 1.

    Supplementary Dataset

    SILAC (Supplementary_Dataset_SILAC)