from bench to cageside: immunology of humanizing...
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From Bench to Cageside:Immunology of Humanizing Mice
Zachary T Freeman
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Goals• Understand relevant immunology related to
humanized mice• Understand three main components that contribute to
formation of humanized mice• Clinical management of severely immunodeficient
mice
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Goals• Understand relevant immunology related to
humanized mice• Understand three main components that contribute to
formation of humanized mice• Clinical management of severely immunodeficient
mice
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Dranoff, G Nat. Rev. Cancer, 2004
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Innate Lymphoid Cells (ILCs) Are a Novel Cell Type for Immune Responses
Shortman K, et al Nat Rev Imm2002
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Innate Lymphoid Cells (ILCs) Are a Novel Cell Type for Immune Responses
Spits H, Nat Rev Imm, 2013
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Humoral Immunity Cellular Immunity
Kumar et al. Robbins Basic Pathology, 8 edn. 2007, pp. 109, Copyright Elsevier.
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B Cells Produced Antibodies as the Main Mediator of Humoral Immunity
Cooper MD, Nat Rev Imm, 2015
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Zou W, Nat Rev Imm, 2010
CD4 T Cells Coordinate and Modulate Cellular Immune Responses
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Restifo NP, Nat Rev Imm, 2012
CD8 T Cells Mediate Effector Portion of Cellular Immunity
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Goals• Understand relevant immunology related to
humanized mice• Understand three main components that contribute to
formation of humanized mice• Clinical management of severely immunodeficient
mice
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Crotty S, Nat Rev Imm, 2015
T and B cell Interactions are Important for Protective Immunity
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https://www.jax.org/jax-mice-and-services/in-vivo-pharmacology/humanized-mice/cd34
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What is an NSG mouse?
https://www.jax.org/strain/005557
NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ
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Non Obese Diabetic Mice Have Altered Innate and Adaptive Immunophenotype
Calis JJA, Trends in Imm, 2014
• T cell mediated islet destruction/diabetes• Defective NK cell function• Defective macrophage cytokine production • Altered immunoregulatory functions of antigen
presenting cells
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What is an NSG mouse?
https://www.jax.org/strain/005557
NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ
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Antigen Specificity Defines B and T cells
Calis JJA, Trends in Imm, 2014
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B and T cell Receptors Share Common Features
Calis JJA, Trends in Imm, 2014
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Lymphoid Specific General
Villartay, JP, Nat Rev Imm, 2003
Deletion of Prkdc Leads to Defect in DNA Repair Mechanisms in All Cells
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What is an NSG mouse?
https://www.jax.org/strain/005557
NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ
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IL-2 Receptor Gamma is Essential B, T, NK and DC Function
Rochman Y, Nat Rev Imm, 2009
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https://www.jax.org/jax-mice-and-services/in-vivo-pharmacology/humanized-mice/cd34
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Ionizing Radiation Has Variable Effects on Immune Cells
Finkelstein SE, et al Frontiers in Oncology, 2012
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https://www.jax.org/jax-mice-and-services/in-vivo-pharmacology/humanized-mice/cd34
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Origins of “Humanizing” Component
• Peripheral blood mononuclear cells (PBMC/PBL)• CD34+ hematopoietic stem cells (HSCs)• Bone marrow, liver, thymus (BLT mice)
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CD34+ Cells Serve as Total Precursor to Immune System
Sharpless NE et al, Nat Rev Mol Cell Bio, 2007
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Shultz LD, Nat Rev Imm, 2012
Unknown Cross talk between Mouse and Human Cytokines
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• S: Human Stem Cell Factor (SCF) gene• maintains normal basal hematopoiesis.
• GM: Human Granulocyte/Macrophage-colony stimulating factor 2 (GM-CSF)• Innate cell numbers and function
• 3: Human interleukin-3 (IL-3) • multipotent hematopoietic stem cells
differentiate into myeloid progenitor cells• myeloid lineage to proliferate
Human Cytokines Enhance Innate Cell Engraftment in NSG-SGM3 Mice
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Shultz LD, Nat Rev Imm, 2012
Human Cytokines Enhance Innate Cell Engraftment in Humanized Mice
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Goals• Understand relevant immunology related to
humanized mice• Understand three main components that contribute to
formation of humanized mice• Clinical management of severely immunodeficient
mice
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Immunology Problems Underlying Clinical Disease in Humanized Mice
• Increased susceptibility to commensal/opportunisitcorganisms• Cornybacterium bovis• Klebsiella oxytoca• Pasturella pneumonia• Pseudomonas sp• Staphlococcus sp• Pneumocystis murina
• Graft Versus Host Disease• Limited resources for the immune system• Risk of anthropozoonosis
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Human Immune Cells Do No Recognize Murine Immune Cells
Kobayashi KS, Nature Reviews Immunology, 2012
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Human Immune Cells Do No Recognize Murine Immune Cells
Bleakley M, Nature Reviews Cancer, 2004
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Limited Host Resources Alter Immune Cell Expansion and Homeostasis
Badovinac VP, Immunity 2007
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Unknown Risk of Anthropozoonosis?
• Human immune cells host to many infectious pathogens• HIV, EBV, CMV, HTLV
• Unknown risk of infection for severely immunodeficient mouse
• Unclear if murine immune system responds to these organisms
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Management of Humanized Mice• Barrier housing
• Air locks/showers• Designated clothing• PPE• PAPRs
• Individually ventilated cages• HEPA filtered air• Automated water
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Management of Humanized Mice• Vary levels of “sterilization”
• Cages• Food• Bedding• Laminar flow hoods
• Food/water practices• Acidification of water• Prophylactic antibiotics not recommended
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Management of Humanized Mice• Infectious Agent Surveillance
• Sentinel selection• Age?• Immune status?• Direct vs. indirect
• Agent exclusion• Testing methodology/frequency
• Limited information about efficacy of treatment• Rederivation to clean up infections
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Management of Humanized Mice
https://www.jax.org/news-and-insights/jax-blog/2015/april/top-5-tips-to-take-care-of-your-nsg-mice
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Basic immunology of humanizing mice is important for clinical management and use of the model.
Questions?
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Acknowledgements
• Jessica Xu DVM• ULAM faculty and residents
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References• 1.Badovinac, V.P., Haring, J.S. & Harty, J.T. Initial T cell receptor transgenic cell precursor frequency dictates critical aspects of the
CD8(+) T cell response to infection. Immunity 26, 827-841 (2007).• 2.Bleakley, M. & Riddell, S.R. Molecules and mechanisms of the graft-versus-leukaemia effect. Nat Rev Cancer 4, 371-380 (2004).• 3.Calis, J.J. & Rosenberg, B.R. Characterizing immune repertoires by high throughput sequencing: strategies and applications. Trends
Immunol 35, 581-590 (2014).• 4.Cooper, M.D. The early history of B cells. Nat Rev Immunol 15, 191-197 (2015).• 5.Crotty, S. A brief history of T cell help to B cells. Nat Rev Immunol 15, 185-189 (2015).• 6.de Villartay, J.P., Fischer, A. & Durandy, A. The mechanisms of immune diversification and their disorders. Nat Rev Immunol 3, 962-
972 (2003).• 7.Dranoff, G. Cytokines in cancer pathogenesis and cancer therapy. Nat Rev Cancer 4, 11-22 (2004).• 8. Finkelstein, S.E. & Fishman, M. Clinical opportunities in combining immunotherapy with radiation therapy. Front Oncol 2, 169
(2012).• 9. Kobayashi, K.S. & van den Elsen, P.J. NLRC5: a key regulator of MHC class I-dependent immune responses. Nat Rev Immunol 12,
813-820 (2012).• 10. Restifo, N.P., Dudley, M.E. & Rosenberg, S.A. Adoptive immunotherapy for cancer: harnessing the T cell response. Nat Rev
Immunol 12, 269-281 (2012).• 11. Rochman, Y., Spolski, R. & Leonard, W.J. New insights into the regulation of T cells by gamma(c) family cytokines. Nat Rev
Immunol 9, 480-490 (2009).• 12. Sharpless, N.E. & DePinho, R.A. How stem cells age and why this makes us grow old. Nat Rev Mol Cell Biol 8, 703-713 (2007).• 13. Shortman, K. & Liu, Y.J. Mouse and human dendritic cell subtypes. Nat Rev Immunol 2, 151-161 (2002).• 14. Spits, H. et al. Innate lymphoid cells--a proposal for uniform nomenclature. Nat Rev Immunol 13, 145-149 (2013).• 15. Walsh, N.C. et al. Humanized Mouse Models of Clinical Disease. Annu Rev Pathol 12, 187-215 (2017).• 16. Zou, W. & Restifo, N.P. T(H)17 cells in tumour immunity and immunotherapy. Nat Rev Immunol 10, 248-256 (2010).• 17. Kumar et al. Robbins Basic Pathology, 8 edn. 2007, pp. 109, Copyright Elsevier.• https://www.jax.org/strain/005557• https://www.jax.org/news-and-insights/jax-blog/2015/april/top-5-tips-to-take-care-of-your-nsg-mice• https://www.jax.org/jax-mice-and-services/in-vivo-pharmacology/humanized-mice/cd34