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Humanized mice for immune system investigation: progress, promise and challenges

Key Points

  • Severely immunodeficient mice engrafted with functional human cells and tissues, known as 'humanized' mice, facilitate progress in studies of human haematopoiesis, immunity, gene therapy, infectious diseases, cancer and regenerative medicine.

  • Mice homozygous for the severe combined immunodeficiency (scid) gene mutation or for targeted mutations at the recombination-activating gene 1 (Rag1) or Rag2 loci, that also have a targeted mutation at the interleukin-2 receptor γ-chain (Il2rg) locus, support high levels of engraftment and function of human haematopoietic stem cells (HSCs) and human immune systems.

  • Advances in humanized mice over the past few years have included approaches to decrease host innate immune responses. In addition, humanized mouse models have benefited greatly from the identification of human species-specific molecules that are crucial for the engraftment and function of human haematopoietic and immune systems and the expression of these molecules in the immunodeficient recipient.

  • The development of humanized mice with functional human immune systems (generated by the engraftment of human lymphoid tissues, HSCs or peripheral blood mononuclear cells) provides an opportunity to carry out translational research on human immunity and autoimmune diseases, and for the study of the biology of the human pathogens responsible for AIDS and several other human-specific infectious diseases.

  • Humanized mice are being used as hosts for primary human tumours for studies of tumour growth and metastasis and for experimental cancer therapy. The phenotypical and functional characterization of human tumour stem cells is also being advanced through the study of humanized mice.

  • The potential for new advances in our understanding of human immunology and other areas of human biology that are supported by studies in humanized mice remains promising. Additional genetic and technological modifications continue to accelerate progress towards the development of a robust functional human immune system in humanized mice.

Abstract

Significant advances in our understanding of the in vivo functions of human cells and tissues and the human immune system have resulted from the development of 'humanized' mouse strains that are based on severely immunodeficient mice with mutations in the interleukin-2 receptor common γ-chain locus. These mouse strains support the engraftment of a functional human immune system and permit detailed analyses of human immune biology, development and functions. In this Review, we discuss recent advances in the development and utilization of humanized mice, the lessons learnt, the remaining challenges and the promise of using humanized mice for the in vivo study of human immunology.

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Figure 1: Approaches for the engraftment of human immune systems in immunodeficient mice.
Figure 2: Cytokines expressed transgenically in immunodeficient Il2rgnull strains.

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Acknowledgements

We thank D. Serreze and R. Maser for critical review of the manuscript. The authors are supported by the US National Institutes of Health (grants AI46629, UO1 DK089572, DK092758 and AI073146), an institutional Diabetes Endocrinology Research Center grant (DK32520), a Cancer Core Grant (CA034196), the Juvenile Diabetes Research Foundation International, and the Leona M. and Harry B. Helmsley Charitable Trust. The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of the US National Institutes of Health.

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Correspondence to Leonard D. Shultz.

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Leonard D. Shultz, Michael A. Brehm and Dale L. Greiner are consultants for ViaCord Inc.

J. Victor Garcia-Martinez is a consultant for Calimmune, GlaxoSmithKline and Boehringer Ingelheim.

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Glossary

γc

A type I cytokine receptor chain that is shared by the receptors for the interleukins IL-2, IL-4, IL-7, IL-9, IL-15 and IL-21.

Prkdc scid

(Often abbreviated as scid). A mutant allele that causes a defect in double-strand DNA break repair that prevents B cell receptor and T cell receptor recombination. Mice with this mutation lack functional B and T cells.

Lentiviral vectors

Vectors that are based on slowly replicating retroviruses. They have a more complex genomic structure than oncoretroviruses (which require cell division for stable integration into the genome of a cell), and they express several accessory proteins in addition to Gag, Pol and Env. The main advantage of using these vectors for gene therapy is their relatively high efficiency of stably transducing quiescent cells.

Bacterial artificial chromosomes

(BACs). Cloning vectors derived from single-copy F-plasmids of Escherichia coli that carry the F replication and partitioning systems that ensure low copy number and the faithful segregation of plasmid DNA into daughter cells. Large genomic fragments can be cloned into such vectors and they are faithfully replicated, which makes BACs useful for constructing genomic libraries.

Knock-in technology

The introduction of a transgene into a precise location in the genome, rather than a random integration site. Knocking-in uses the same technique of homologous recombination as a knockout strategy, but the targeting vector is designed to allow expression of the introduced transgene under the control of the regulatory elements of the targeted gene.

Thrombocytopenia

A reduced number of circulating platelets, owing either to the failure of production from bone-marrow megakaryocytes or to increased clearance from the circulation, predominantly in the spleen.

Graft-versus-host disease

(GVHD). Tissue damage in a recipient of allogeneic or xenogeneic tissue (usually a bone marrow transplant) that results from the activity of donor cytotoxic T lymphocytes that recognize the tissues of the recipient as foreign. GVHD varies markedly in extent, but it can be life-threatening in severe cases. Damage to the liver, skin and gut mucosa are common clinical manifestations.

Severe combined immunodeficiency

(SCID). Humans or mice with this rare genetic disorder lack functional B and T cells owing to a mutation in a gene that is involved in B cell and T cell development, and consequently they suffer from recurrent infections. Several forms of SCID have been described, including those caused by defects in the IL-2 receptor γ-chain (which is shared by several interleukin receptors), in PRKDC (DNA-dependent protein kinase catalytic subunit), in Janus kinase 3 and in adenosine deaminase.

Central memory T cells

Antigen-experienced T cells that lack immediate effector function but can mediate rapid recall responses. They also rapidly develop the phenotype and function of effector memory T cells after re-stimulation with antigen. Central memory T cells retain the migratory properties of naive T cells and therefore circulate through the secondary lymphoid organs.

Effector memory T cells

Memory T cells that home to peripheral tissues. They are responsible for immediate protection following re-infection.

Germinal centres

Lymphoid structures that arise within follicles after immunization with, or exposure to, a T cell-dependent antigen. They are specialized for facilitating the development of high-affinity, long-lived plasma cells and memory B cells.

Delayed-type hypersensitivity

A cellular immune response to an antigen that develops over 24–72 hours, involves the infiltration of T cells and monocytes, and is dependent on the production of T helper 1 cell-specific cytokines.

α-galactosylceramide

A synthetic or marine-sponge-derived glycolipid containing an α-anomeric glycosidic linkage of the galactose residue to the sphingosine base. This lipid, and structurally related ones, potently activates CD1d-restricted natural killer T cells that express the semi-invariant Vα214–Jα18 T cell receptor in mice (or the Vα24–Jα18 equivalent receptor in humans).

Systemic lupus erythematosus

(SLE). An autoimmune disease in which autoantibodies specific for DNA, RNA or proteins associated with nucleic acids form immune complexes. These complexes damage small blood vessels, especially in the kidneys. Patients with SLE generally have abnormal B and T cell function.

Lymphoid tissue inducer cell

A cell that is present in developing lymph nodes, Peyer's patches and nasopharynx-associated lymphoid tissue (NALT). Lymphoid tissue inducer cells are required for the development of these lymphoid organs. The inductive capacity of these cells for the generation of Peyer's patches and NALT has been shown by adoptive transfer, and it is generally assumed that they have a similar function in the formation of lymph nodes.

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Shultz, L., Brehm, M., Garcia-Martinez, J. et al. Humanized mice for immune system investigation: progress, promise and challenges. Nat Rev Immunol 12, 786–798 (2012). https://doi.org/10.1038/nri3311

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