Skip to main content

Characterization of Immune Cell Subset Expansion in Response to Therapeutic Treatment in Mice

  • Protocol
  • First Online:
T-Cell Receptor Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2111))

Abstract

Flow cytometry has revolutionized the field of molecular immunology, enabling the monitoring and characterization of immune events at the single-cell level. Here, we describe a flow cytometry-based workflow to quantify the activation of specific immune cell subsets in mice in response to a molecular intervention. Compared to laborious long-term disease models, this technique allows for relatively rapid evaluation of candidate therapeutics designed to elicit a targeted immune response. This approach has the range to address both disease applications in which an immunostimulatory effect would be desired (e.g., cancer, infectious disease) or those in which an immunosuppressive effect would be desired (e.g., autoimmune disorders, transplantation medicine). Overall, our technique presents a powerful and accessible strategy for preliminary in vivo assessment of potential immunotherapeutics.

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 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 149.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
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. Muirhead KA, Horan PK, Poste G (1985) Flow cytometry: present and future. Nat Biotechnol 3(4):337–356

    Article  CAS  Google Scholar 

  2. Adan A, Alizada G, Kiraz Y, Baran Y, Nalbant A (2017) Flow cytometry: basic principles and applications. Crit Rev Biotechnol 37(2):163–176

    Article  CAS  Google Scholar 

  3. Macey MG (ed) (2007) Flow cytometry: principles and applications [Internet]. Humana: Totowa, NJ. http://www.myilibrary.com?id=97216. Accessed 3 Apr 2019

  4. Spangler JB, Moraga I, Mendoza JL, Garcia KC (2015) Insights into cytokine-receptor interactions from cytokine engineering. Annu Rev Immunol 33(1):139–167

    Article  CAS  Google Scholar 

  5. Kureshi R, Bahri M, Spangler JB (2018) Reprogramming immune proteins as therapeutics using molecular engineering. Curr Opin Chem Eng 19:27–34

    Article  Google Scholar 

  6. Taebel DW (1990) The importance of animals in biomedical research. Wis Med J 89(4):155. 158

    CAS  PubMed  Google Scholar 

  7. Rosenberg SA, Restifo NP, Yang JC, Morgan RA, Dudley ME (2008) Adoptive cell transfer: a clinical path to effective cancer immunotherapy. Nat Rev Cancer 8(4):299–308

    Article  CAS  Google Scholar 

  8. Ford ML, Koehn BH, Wagener ME, Jiang W, Gangappa S, Pearson TC and Larsen CP (2007) Antigen-specific precursor frequency impacts T cell proliferation, differentiation, and requirement for costimulation. J Exp Med 204(2):299–309. 

    Article  CAS  Google Scholar 

  9. Svedova M, Masin J, Fiser R, Cerny O, Tomala J, Freudenberg M, Tuckova L, Kovar M, Dadaglio G, Adkins I, Sebo P (2016) Pore-formation by adenylate cyclase toxoid activates dendritic cells to prime CD8+ and CD4+ T cells. Immunol Cell Biol 94(4):322–333

    Article  CAS  Google Scholar 

  10. Skopova K, Tomalova B, Kanchev I, Rossmann P, Svedova M, Adkins I, Bibova I, Tomala J, Masin J, Guiso N, Osicka R, Sedlacek R, Kovar M, Sebo P (2017) Cyclic AMP-elevating capacity of adenylate cyclase toxin-hemolysin is sufficient for lung infection but not for full virulence of Bordetella pertussis. Infect Immun 85(6):e00937

    Article  Google Scholar 

  11. Spangler JB, Tomala J, Luca VC, Jude KM, Dong S, Ring AM, Votavova P, Pepper M, Kovar M, Garcia KC (2015) Antibodies to interleukin-2 elicit selective T cell subset potentiation through distinct conformational mechanisms. Immunity 42(5):815–825

    Article  CAS  Google Scholar 

  12. Spangler JB, Trotta E, Tomala J, Peck A, Young TA, Savvides CS, Silveria S, Votavova P, Salafsky J, Pande VS, Kovar M, Bluestone JA, Garcia KC (2018) Engineering a single-agent cytokine/antibody fusion that selectively expands regulatory T cells for autoimmune disease therapy. J Immunol 201(7):2094–2106

    Article  CAS  Google Scholar 

  13. Trotta E, Bessette PH, Silveria SL, Ely LK, Jude KM, Le DT, Holst CR, Coyle A, Potempa M, Lanier LL, Garcia KC, Crellin NK, Rondon IJ, Bluestone JA (2018) A human anti-IL-2 antibody that potentiates regulatory T cells by a structure-based mechanism. Nat Med 24(7):1005–1014

    Article  CAS  Google Scholar 

  14. Sockolosky JT, Trotta E, Parisi G, Picton L, Su LL, Le AC, Chhabra A, Silveria SL, George BM, King IC, Tiffany MR, Jude K, Sibener LV, Baker D, Shizuru JA, Ribas A, Bluestone JA, Garcia KC (2018) Selective targeting of engineered T cells using orthogonal IL-2 cytokine-receptor complexes. Science 359(6379):1037–1042

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jamie B. Spangler .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2020 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Tomala, J., Spangler, J.B. (2020). Characterization of Immune Cell Subset Expansion in Response to Therapeutic Treatment in Mice. In: Liu, C. (eds) T-Cell Receptor Signaling. Methods in Molecular Biology, vol 2111. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0266-9_9

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-0266-9_9

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0265-2

  • Online ISBN: 978-1-0716-0266-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics