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.
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References
Muirhead KA, Horan PK, Poste G (1985) Flow cytometry: present and future. Nat Biotechnol 3(4):337–356
Adan A, Alizada G, Kiraz Y, Baran Y, Nalbant A (2017) Flow cytometry: basic principles and applications. Crit Rev Biotechnol 37(2):163–176
Macey MG (ed) (2007) Flow cytometry: principles and applications [Internet]. Humana: Totowa, NJ. http://www.myilibrary.com?id=97216. Accessed 3 Apr 2019
Spangler JB, Moraga I, Mendoza JL, Garcia KC (2015) Insights into cytokine-receptor interactions from cytokine engineering. Annu Rev Immunol 33(1):139–167
Kureshi R, Bahri M, Spangler JB (2018) Reprogramming immune proteins as therapeutics using molecular engineering. Curr Opin Chem Eng 19:27–34
Taebel DW (1990) The importance of animals in biomedical research. Wis Med J 89(4):155. 158
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
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.Â
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
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
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
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
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
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
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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
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DOI: https://doi.org/10.1007/978-1-0716-0266-9_9
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