Characterization of Immune Cell Subset Expansion in Response to Therapeutic Treatment in Mice
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.
Key wordsImmunology T cell Natural killer cell Flow cytometry Immunophenotyping Immunotherapy
- 3.Macey MG (ed) (2007) Flow cytometry: principles and applications [Internet]. Humana: Totowa, NJ. http://www.myilibrary.com?id=97216. Accessed 3 Apr 2019
- 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):e00937CrossRefGoogle 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):2094–2106CrossRefGoogle 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):1037–1042CrossRefGoogle Scholar