Skip to main content

Isolation of Intratumoral Leukocytes of TLR-Stimulated Tumor-Bearing Mice

  • Protocol
  • First Online:
Innate DNA and RNA Recognition

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1169))

Abstract

Toll-like receptor (TLR) ligands hold promise for cancer immunotherapy. The isolation of intratumoral leukocytes of tumor-bearing mice is a useful technique for analyzing the immunological effects of TLR ligands on the tumor microenvironment. These isolated immune cells can be directly used for analysis (e.g., by flow cytometry) or cultured for functional in vitro studies. Here, we describe the isolation of intratumoral leukocytes by density gradient centrifugation. This technique can be used to isolate leukocytes from freshly dissected murine tumors.

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 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
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. Kanzler H, Barrat FJ, Hessel EM, Coffman RL (2007) Therapeutic targeting of innate immunity with Toll-like receptor agonists and antagonists. Nat Med 13(5):552–559

    Article  CAS  PubMed  Google Scholar 

  2. Heckelsmiller K, Rall K, Beck S, Schlamp A, Seiderer J, JahrsdoÅNrfer B, Krug A, Rothenfusser S, Endres S, Hartmann G (2002) Peritumoral CpG DNA elicits a coordinated response of CD8 T cells and innate effectors to cure established tumors in a murine colon carcinoma model. J Immunol 169(7):3892–3899

    Article  CAS  PubMed  Google Scholar 

  3. Houot R, Levy R (2009) T-cell modulation combined with intratumoral CpG cures lymphoma in a mouse model without the need for chemotherapy. Blood 113(15):3546–3552

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  4. Krieg AM (2008) Toll-like receptor 9 (TLR9) agonists in the treatment of cancer. Oncogene 27:161–167

    Article  CAS  PubMed  Google Scholar 

  5. Weber JS, Zarour H, Redman B, Trefzer U, O’Day S, van den Eertwegh AJ, Marshall E, Wagner S (2009) Randomized phase 2/3 trial of CpG oligodeoxynucleotide PF-3512676 alone or with dacarbazine for patients with unresectable stage III and IV melanoma. Cancer 115(17):3944–3954

    Article  CAS  PubMed  Google Scholar 

  6. Vollmer J, Krieg AM (2009) Immunotherapeutic applications of CpG oligodeoxynucleotide TLR9 agonists. Adv Drug Deliv Rev 61:195–204

    Article  CAS  PubMed  Google Scholar 

  7. Chang YC, Madkan V, Cook-Norris R, Sra K, Tyring S (2005) Current and potential uses of imiquimod. South Med J 98(9):914–920

    Article  PubMed  Google Scholar 

  8. van Seters M, van Beurden M, ten Kate FJ, Beckmann I, Ewing PC, Eijkemans MJ, Kagie MJ, Meijer CJ, Aaronson NK, Kleinjan A, Heijmans-Antonissen C, Zijlstra FJ, Burger MP, Helmerhorst TJ (2008) Treatment of vulvar intraepithelial neoplasia with topical imiquimod. N Engl J Med 358(14):1465–1473

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

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

Rapp, M., Anz, D., Schnurr, M. (2014). Isolation of Intratumoral Leukocytes of TLR-Stimulated Tumor-Bearing Mice. In: Anders, HJ., Migliorini, A. (eds) Innate DNA and RNA Recognition. Methods in Molecular Biology, vol 1169. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0882-0_16

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-0882-0_16

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0881-3

  • Online ISBN: 978-1-4939-0882-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics