Skip to main content

Experimental Model for Studying the Involvement of Regulatory Cytotoxic T Cells in Bone Resorption

  • Protocol
  • First Online:
Cytotoxic T-Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1186))

Abstract

T cells may affect the activity of osteoclasts, the bone resorbing cells. In particular, recently authors focalize their attention on CD8+ T cells, demonstrating that murine pre-osteoclasts can recruit naïve CD8+ T cells and induce them to differentiate in regulatory cytotoxic T cells (TcREG), which in turn may suppress osteoclast formation and activity. Thus, here we describe the methodological approach needed to obtain TcREG and to evaluate TcREG effects on murine osteoclast activity and formation in an in vitro experimental model.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Mori G, D’Amelio P, Faccio R, Brunetti G (2013) The interplay between the bone and the immune system. Clin Dev Immunol 2013(720504):1–9

    Article  Google Scholar 

  2. Boyle WJ, Simonet WS, Lacey DL (2003) Osteoclast differentiation and activation. Nature 423:337–342

    Article  CAS  PubMed  Google Scholar 

  3. Kong YY, Feige U, Sarosi I, Bolon B, Tafuri A, Morony S, Capparelli C, Li J, Elliott R, McCabe S, Wong T, Campagnuolo G, Moran E, Bogoch ER, Van G, Nguyen LT, Ohashi PS, Lacey DL, Fish E, Boyle WJ, Penninger JM (1999) Activated T cells regulate bone loss and joint destruction in adjuvant arthritis through osteoprotegerin ligand. Nature 402:304–309

    Article  CAS  PubMed  Google Scholar 

  4. Colucci S, Brunetti G, Cantatore FP, Oranger A, Mori G, Quarta L, Cirulli N, Mancini L, Corrado A, Grassi FR, Grano M (2007) Lymphocytes and synovial fluid fibroblasts support osteoclastogenesis through RANKL, TNFalpha, and IL-7 in an in vitro model derived from human psoriatic arthritis. J Pathol 212:47–55

    Article  CAS  PubMed  Google Scholar 

  5. Teng YT, Nguyen H, Gao X, Kong YY, Gorczynski RM, Singh B, Ellen RP, Penninger JM (2000) Functional human T-cell immunity and osteoprotegerin ligand control alveolar bone destruction in periodontal infection. J Clin Invest 106:R59–R67

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  6. Brunetti G, Colucci S, Pignataro P, Coricciati M, Mori G, Cirulli N, Zallone A, Grassi FR, Grano M (2005) T cells support osteoclastogenesis in an in vitro model derived from human periodontitis patients. J Periodontol 76:1675–1680

    Article  CAS  PubMed  Google Scholar 

  7. Colucci S, Brunetti G, Rizzi R, Zonno A, Mori G, Colaianni G, Del Prete D, Faccio R, Liso A, Capalbo S, Liso V, Zallone A, Grano M (2004) T cells support osteoclastogenesis in an in vitro model derived from human multiple myeloma bone disease: the role of the OPG/TRAIL interaction. Blood 104:3722–3730

    Article  CAS  PubMed  Google Scholar 

  8. Roato I, Grano M, Brunetti G, Colucci S, Mussa A, Bertetto O, Ferracini R (2005) Mechanisms of spontaneous osteoclastogenesis in cancer with bone involvement. FASEB J 19:228–230

    CAS  PubMed  Google Scholar 

  9. Faienza MF, Brunetti G, Colucci S, Piacente L, Ciccarelli M, Giordani L, Del Vecchio GC, D’Amore M, Albanese L, Cavallo L, Grano M (2009) Osteoclastogenesis in children with 21-hydroxylase deficiency on long term glucocorticoid therapy: the role of RANKL/OPG imbalance. J Clin Endocrinol Metab 94:2269–2276

    Article  CAS  PubMed  Google Scholar 

  10. Choi Y, Woo KM, Ko SH, Lee YJ, Park SJ, Kim HM, Kwon BS (2001) Osteoclastogenesis is enhanced by activated B cells but suppressed by activated CD8(+) T cells. Eur J Immunol 31:2179–2188

    Article  CAS  PubMed  Google Scholar 

  11. Fontenot JD, Gavin MA, Rudensky AY (2003) Foxp3 programs the development and function of CD4 + CD25+ regulatory T cells. Nat Immunol 4:330–336

    Article  CAS  PubMed  Google Scholar 

  12. Hori S, Nomura T, Sakaguchi S (2003) Control of regulatory T cell development by the transcription factor Foxp3. Science 299:1057–1061

    Article  CAS  PubMed  Google Scholar 

  13. Zaiss MM, Axmann R, Zwerina J, Polzer K, Gückel E, Skapenko A, Schulze-Koops H, Horwood N, Cope A, Schett G (2007) Treg cells suppress osteoclast formation: a new link between the immune system and bone. Arthritis Rheum 56:4104–4112

    Article  CAS  PubMed  Google Scholar 

  14. Buchwald ZS, Kiesel JR, DiPaolo R, Pagadala MS, Aurora R (2012) Osteoclast activated FoxP3+ CD8+ T-cells suppress bone resorption in vitro. PLoS One 7:e38199

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  15. Sharabi A, Mozes E (2008) The suppression of murine lupus by a tolerogenic peptide involves foxp3-expressing CD8 cells that are required for the optimal induction and function of foxp3-expressing CD4 cells. J Immunol 181:3243–3251

    Article  CAS  PubMed  Google Scholar 

  16. Kiniwa Y, Miyahara Y, Wang HY, Peng W, Peng G, Wheeler TM, Thompson TC, Old LJ, Wang RF (2007) CD8+ Foxp3+ regulatory T cells mediate immunosuppression in prostate cancer. Clin Cancer Res 13:6947–6958

    Article  CAS  PubMed  Google Scholar 

  17. Singh RP, La Cava A, Wong M, Ebling F, Hahn BH (2007) CD8+ T cell mediated suppression of autoimmunity in a murine lupus model of peptide induced immune tolerance depends on Foxp3 expression. J Immunol 178:7649–7657

    Article  CAS  PubMed  Google Scholar 

  18. Kiesel J, Miller C, Abu-Amer Y, Aurora R (2007) Systems level analysis of osteoclastogenesis reveals intrinsic and extrinsic regulatory interactions. Dev Dyn 236:2181–2197

    Article  CAS  PubMed  Google Scholar 

  19. Kiesel JR, Buchwald ZS, Aurora R (2009) Cross-presentation by osteoclasts induces FoxP3 in CD8+ T cells. J Immunol 182:5477–5487

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Giacomina Brunetti Ph.D. .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media New York

About this protocol

Cite this protocol

Brunetti, G., Oranger, A., Colucci, S., Grano, M. (2014). Experimental Model for Studying the Involvement of Regulatory Cytotoxic T Cells in Bone Resorption. In: Ranieri, E. (eds) Cytotoxic T-Cells. Methods in Molecular Biology, vol 1186. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1158-5_15

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-1158-5_15

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1157-8

  • Online ISBN: 978-1-4939-1158-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics