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Humanized Mice as Unique Tools for Human-Specific Studies

  • Kylie Su Mei Yong
  • Zhisheng Her
  • Qingfeng ChenEmail author
Open Access
Review

Abstract

With an increasing human population, medical research is pushed to progress into an era of precision therapy. Humanized mice are at the very heart of this new forefront where it is acutely required to decipher human-specific disease pathogenesis and test an array of novel therapeutics. In this review, “humanized” mice are defined as immunodeficient mouse engrafted with functional human biological systems. Over the past decade, researchers have been conscientiously making improvements on the development of humanized mice as a model to closely recapitulate disease pathogenesis and drug mechanisms in humans. Currently, literature is rife with descriptions of novel and innovative humanized mouse models that hold a significant promise to become a panacea for drug innovations to treat and control conditions such as infectious disease and cancer. This review will focus on the background of humanized mice, diseases, and human-specific therapeutics tested on this platform as well as solutions to improve humanized mice for future clinical use.

Keywords

Humanized mice Human specificity Precision therapy Human diseases Drug testing 

Introduction

Fundamental understandings of many biological processes that occur in humans have evolved from experimental studies on animal models, particularly non-human rodents and non-human primates (NHPs) (Hatziioannou and Evans 2012; Phillips et al. 2014). A major technical barrier in translating these discoveries to treatments is caused by differences in the biological systems between animals and humans (Greek and Rice 2012; Mestas and Hughes 2004; Shanks et al. 2009; Van der Worp et al. 2010). For example, functional Toll-like receptor 10 (TLR10) is absent in mice (Oosting et al. 2014) and cell expression marker CD28 is expressed on 100% of CD4+ and CD8+ T cells in mice but only on 80% of CD4+ and 50% CD8+ T cells in humans (Beyersdorf et al. 2015). Due to these differences, it is common that animal models are refractory to many infectious (Bäumler and Fang 2013; Carlton et al. 2008; Fauci 1988; Pain et al. 2008; Ploss et al. 2009), therapeutic (McKenzie et al. 1995; Rehman et al. 2011), or immunomodulatory agents (Attarwala 2010; Tsoneva et al. 2017) that are human-specific.

To address the limitations of translating discoveries on non-human animal models to clinical applications, a platform known as “humanized mice” was engineered to simulate humans at a cellular and molecular level (Bosma et al. 1983; Pearson et al. 2008). Humanized mice generated in recent years encompass functional human immune systems with expansive capabilities (Rongvaux et al. 2014) and are unprecedented platforms used for understanding disease pathogenesis and evaluation of compounds to treat a variety of human diseases which include but are not limited to, cancer (Her et al. 2017; Ito et al. 2009; Miyakawa et al. 2004; Pan et al. 2017), infectious disease (Amaladoss et al. 2015; Frias-Staheli et al. 2014; Keng et al. 2015; Yajima et al. 2008), autoimmune disease (Gunawan et al. 2017; Viehmann Milam et al. 2014; Young et al. 2015; Zayoud et al. 2013), and graft-versus-host disease (GvHD) (King et al. 2008; Kirkiles-Smith et al. 2009; Tobin et al. 2013; Zhao et al. 2015).

This review covers the background of humanized mice, diseases modelled on these platforms, human-specific therapeutics tested, and suggestions for overcoming remaining challenges to improve humanized mouse models for clinical applications.

Evolving History of Humanized Mice

There has been a constant pursuit to engineer novel immunodeficient mouse models via gene deletion or backcrossing strains with mutations in essential molecular compartments such as, T cells, B cells, macrophages, natural killer (NK) cells, cytokines, TLRs, and transcription factors (Pearson et al. 2008). The aim of introducing these mutations is to reduce murine cells and increase the engraftment of human cells and tissues to better recapitulate human immune responses (Aryee et al. 2014; Billerbeck et al. 2011; Chen et al. 2009; Rongvaux et al. 2014; Yao et al. 2016).

Tracing the roots of humanized mice, the discovery of non-human animal models xenotransplanted with cells and tissues of human origin was credited to the invention of C.B-17-Prkdcscid (CB17-scid) mice (Bosma et al. 1983). Derived from backcrossing C57BL/Ka and BALB/c, this mouse features loss of function mutation in a gene known as protein kinase, DNA-activated, catalytic polypeptide (PRKDC). In normal physiological conditions, PRKDC is essential for resolving breaks in DNA strands during variable, diversity, and joining [V(D)J] recombination for the development of T and B cells (Blunt et al. 1996; Finnie et al. 1996; Lieber et al. 1988; Taccioli et al. 1998). Non-functional PRKDC gene leads to impaired development of T and B cells resulting in syndrome known as severe combined immunodeficiency (scid) (Bosma and Carroll 1991). Despite efforts in creating CB17-scid mice, this model was not used in many experiments due to the poor engraftment of human hematopoietic stem cells (HSCs) (Bosma et al. 1983).

Further research saw the transfer of scid mutation onto a mouse of non-obese diabetic (NOD) background, creating NOD-scid mice which lacked T cells, B cells, and NK cells. This mouse allowed a slightly higher level of human cell reconstitution (Van der Loo et al. 1998). However, the biggest breakthrough in humanized mice only occurred when mutant interleukin 2 receptor α (IL2rα) gene was introduced into NOD-scid mice, creating NOD-scid-γcnull mice (NSG or NOG), which exhibited defective mouse cytokines IL-2, IL-4, IL-7, IL-9, and IL-15 (Ishikawa et al. 2005; Ito et al. 2002; Shultz et al. 2005). Knock-out of recombination activating gene (RAG) 1 or 2 (RAG1null and RAG2null) caused even greater immunodeficiencies including an absence of NK cells, T cells, B cells, and impaired macrophage and dendritic cell (DC) subsets (Harris and Badowski 2014; Watanabe et al. 2007). However, an absence of human leukocyte antigen (HLA) in these models resulted in engrafted human pre-T cells being “educated” and selected on mouse thymic epithelium and major histocompatibility complexes (MHCs) (Shultz et al. 2010). Due to this limitation, engrafted human T cells were unable to recognise human antigen-presenting cells, and hence, these mice had impaired immunoglobulin (Ig) class switching and disorganised secondary lymphoid structures (Shultz et al. 2010, 2012). To overcome this hurdle, HLA class I and II transgenes were added into NSG mice allowing the development of human T-cell repertoires and responses (Brehm et al. 2013; Shultz et al. 2010).

Improved models of immunodeficient mice enabled an increase in well-differentiated multilineage human hematopoietic cells, high levels of functional human cell reconstitution and an ability to be engrafted with tissues such as thymus, skin, liver, islets, solid tumors, and blood cancers (Ito et al. 2002). These inventions cascaded into a series of immunodeficient mice and their variants (BRG, NOG, NRG) (Ali et al. 2012; Grover et al. 2017; Ishikawa et al. 2005; Katano et al. 2014; Koboziev et al. 2015; Shultz et al. 2005) being innovated which enabled in-depth analysis in research areas, such as human hematopoiesis (Rongvaux et al. 2011; Yong et al. 2016), innate and adaptive immunity (Brehm et al. 2010; Pearson et al. 2008), autoimmunity (Gunawan et al. 2017; Viehmann Milam et al. 2014), infectious disease (Keng et al. 2015; Lüdtke et al. 2015; Wege et al. 2012), cancer biology (Chang et al. 2015; Her et al. 2017; Morton et al. 2016), and GvHD (King et al. 2008; Kirkiles-Smith et al. 2009; Zhao et al. 2015), in-turn, facilitating the development of therapeutic agents and novel vaccines. An overview of genotypic and physiological characteristics of each model is outlined in Tables 1 and 2.

Table 1

Platforms for human immune system engrafted mice

Name

C.B-17-scid

NOD-scid

BRG

NOG

NSG™, NOD-scid-γ

NRG, NOD Rag

Nomenclature

C.B-Igh-1b/IcrTac-Prkdcscid

NOD.CB17-Prkdcscid/J

C.Cg-Rag2tm1Fwa Il2rgtm1Sug/JicTac

NOD.Cg-Prkdcscid Il2rgtm1Sug/JicTac

NOD.Cg-Prkdcscid

Il2rgtm1Wjl/SzJ

NOD.Cg-Rag1tm1Mom

Il2rgtm1Wjl/SzJ

Engraftment method for humanization

HSPCs

BM cells

Spleen cells

HSPCs

PBMCs

Thymus and liver under kidney capsule with matching engraftment of HSPCs from FL

Cancer derived from patients and cell lines

HSPCs

PBMCs

HSPCs

PBMCs

Thymus and liver under kidney capsule with matching engraftment of HSPCs from FL

Cancer derived from patient and cell lines

HSPCs

PBMCs

Thymus and liver under kidney capsule with matching engraftment of HSPCs from FL

Cancer derived from patients and cell lines

HSPCs

PBMCs

Thymus and liver under kidney capsule with matching engraftment of HSPCs from FL

Cancer derived from patients and cell lines

Limitations

Low tolerance for irradiation

Intact innate immune system

Rejection of engraftments

Spontaneous development of thymic lymphomas

Short lifespan

Low tolerance for irradiation

Spontaneous development of thymic lymphomas

Not all cancers can be engrafted

Spontaneous development of thymic lymphomas

Low tolerance for irradiation

Not all cancers can be engrafted

High occurrence of tumor metastasis

Low tolerance for irradiation

Spontaneous development of thymic lymphomas

Not all cancers can be engrafted

Requires a higher dose of irradiation

Not all cancers can be engrafted

Applications

GvHD

Autoimmune type I diabetes

Oncological studies

Immune system

Infectious diseases

Oncological studies

Stem cells

Immune system

Infectious diseases

Oncological studies

Drug tests

Stem cells

Immune system

Infectious diseases

Oncological studies

Drug tests

Stem cells

Immune system

Infectious diseases

Oncological studies

Drug tests

Dendritic cells

Yes

Impaired

Impaired

Impaired

Impaired

Impaired

Macrophages

Yes

Impaired

Impaired

Impaired

Impaired

Impaired

NK cells

Yes

No

No

No

No

No

Mature B cells

No

No

No

No

No

No

Mature T cells

No

No

No

No

No

No

Complement

Yes

No

No

No

No

No

Leakiness

Low

Low

No

No

Low

No

Irradiation tolerance

Low

Low

High

Low

Low

High

Lymphoma incidence

High

High

Low

No

No

Low

Median lifespan

< 12 months

< 10 months

Not determined

> 18 months

> 18 months

Not determined

References

Schneider et al. (1997)

Sheng-Tanner et al. (2000)

Xia et al. (2006)

Bastide et al. (2002)

Brehm et al. (2013)

Traggiai et al. (2004)

Ali et al. (2012)

Akkina (2013)

Watanabe et al. (2009)

Akkina (2013)

Yong et al. (2016)

Her et al. (2017)

Harris et al. (2013)

Shultz et al. (2012)

Maykel et al. (2014)

Table 2

Platforms for human immune system engrafted mice

Name

HuNOG-EXL

NSG-SGM3

NSG-HLA-A2

NSG-Ab DR4

MISTRG

NSGW41

Nomenclature

NOD.Cg-Prkdcscid Il2rgtm1Sug Tg(SV40/HTLV-IL3, CSF2)10-7Jic/JicTac

NOD.Cg-Prkdcscid

Il2rgtm1Wjl Tg(CMV, IL3, CSF2, KITLG)1Eav/MloySzJ

NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(HLA-A2.1)1Enge/SzJ

NOD.Cg-Prkdcscid Il2rgtm1Wjl H2-Ab1tm1Gru Tg(HLA-DRB1)31Dmz/SzJ

C;129S4-Rag2tm1.1FlvCsf1tm1(CSF1)FlvCsf2/Il3tm1.1(CSF2,IL3)FlvThpotm1.1(TPO)FlvIl2rgtm1.1FlvTg(SIRPA)1Flv/J

NOD.Cg-KitW41J Prkdcscid Il2rgtm1Wjl/WaskJ

Engraftment method for humanization

HSPCs

PBMCs

Thymus and liver under kidney capsule with matching engraftment of HSPCs from FL

Cancer derived from patients and cell lines

HSPCs

PBMCs

Thymus and liver under kidney capsule with matching engraftment of HSPCs from FL

Cancer derived from patients and cell lines

HSPCs

PBMCs

PBMCs

HSPCs

Human melanoma cell line (Me290)

HSPCs

Limitations

Not all cancers can be engrafted

Mice with high chimeric ratio develop anemia after engraftment

Human cell engraftment does not last more than five months

Low tolerance for irradiation

Low CD45+ human cell engraftment compared to NSG mice

Short lifespan post-engraftment (~ 10–12 weeks) but may be prolonged by avoiding irradiation, using less potent and lower number of stem cells

Not reported

Applications

Stem cells

Immune system

Infectious diseases

Oncological studies

Drug tests

Stem cells

Immune system

Infectious diseases

Oncological studies

Drug tests

Immune system

Oncological studies

Vaccine development

GvHD

Stem cells

Immune system

Oncological studies

Stem cells

Dendritic cells

Impaired

Impaired

Impaired

Impaired

Impaired

Impaired

Macrophages

Impaired

Impaired

Impaired

Impaired

Impaired

Impaired

NK cells

No

No

No

No

No

No

Mature B cells

No

No

No

No

No

No

Mature T cells

No

No

No

No

No

No

Complement

No

No

No

No

No

No

Leakiness

No

No

Low

Low

No

No

Irradiation tolerance

Not determined

Not determined

Low

High

Low

Low

Lymphoma incidence

Not determined

Not determined

No

No

Not determined

Not determined

Median lifespan

> 7 months

> 4 months

> 18 months

Not determined

Not determined

Not determined

References

Fukuchi et al. (1998)

Ito et al. (2013)

Billerbeck et al. (2011)

Whitfield-Larry et al. (2011)

Patton et al. (2015)

Covassin et al. (2011)

Rongvaux et al. (2014)

Rahmig et al. (2016)

HSPCs hematopoietic stem and progenitor cells, FL fetal liver, GvHD graft-versus-host disease, PBMCs peripheral blood mononuclear cells, BM bone marrow

The conventional ways to engraft immunodeficient mice with functional human cells include, intravenous (i.v.) injection of human peripheral blood mononuclear cells (PBMCs) into mice (Hu-PBL-scid) (Duchosal et al. 1992; Harui et al. 2011; King et al. 2008; Tary-Lehmann et al. 1995), injecting CD34+ HSCs obtained from human fetal liver (FL), umbilical cord blood (UBC), bone marrow (BM) or granulocyte-colony-stimulating factor (G-CSF) mobilised peripheral blood (Hu-SRC-scid) (Brehm et al. 2010; Chen et al. 2009, 2012, 2015; Keng et al. 2015; Yong et al. 2016), or i.v. injection of FL HSCs and BM cells paired with transplantation of matching FL and thymus under the kidney capsule to obtain a BM/liver/thymus (BLT) mouse model (Brainard et al. 2009; Covassin et al. 2013; Denton et al. 2008; Lan et al. 2004, 2006; Melkus et al. 2006; Tonomura et al. 2008). Advantages and drawbacks of each method are compared in Table 3. However, despite efforts in optimising humanized mice, critical challenges that remain include: limited fetal samples due to ethical restrictions (Geraghty et al. 2014; Kapp 2006), absence of erythrocytes and neutrophils within reconstituted human immune system (Hu et al. 2011), low and impaired human myeloid cells, dominance of immature B cells (Chen et al. 2012; Lang et al. 2013), and minimal production of antigen-specific IgG class antibodies in humanized mice (Jangalwe et al. 2016).

Table 3

Methods used to establish humanized mouse models

Model

Human PBMCs engrafted into immunodeficient mice

Human HSCs engrafted into immunodeficient mice

Human HSCs, BM, liver, and thymus engrafted into immunodeficient mice

Alternative name

Hu-PBL-scid

Hu-SRC-scid

BLT

Source of cells

Obtained from consented adult donors

FL

UBC

BM

G-CSF mobilised peripheral blood

FL

Fetal BM

Fetal thymus

Method of engraftment

Intravenous injection of mice

Intrahepatic injection of newborn mice within 72 h of birth

Intravenous injection of mice

Implantation of liver and thymus under the kidney capsule

Transplantation of matching HSCs obtained from FL

Advantages

Easy techniques applied

Fast to establish

Presence of functional immune cells such as memory T cells

Excellent in modelling GvHD

Multilineage development of hematopoietic cells

Generation of a naïve immune system

Injection to pups increase human cell reconstitution

Complete and fully functional human immune system

HLA-restricted T cells

Development of a mucosal system similar to humans

Highest level of human cell reconstitution among all the models

Drawbacks

Lack B and myeloid cell engraftment

Engrafted T cells are activated

May develop GvHD

Only suitable for short-term experiments (< 3 months)

Cell differentiation takes a minimum of 10 weeks

Engrafted human T cells are H2 restricted

Contains low levels of human RBCs, polymorphonuclear leukocytes, and megakaryocytes

Time-consuming and difficult as surgical implantation is required

Cell differentiation takes a minimum of 10 weeks

Weak immune responses to xenobiotics

Poor class switching

May develop GvHD

BLT bone marrow/liver/thymus, HSCs hematopoietic stem cells, FL fetal liver, GvHD graft-versus-host disease, PBMCs peripheral blood mononuclear cells, UBC umbilical cord blood, BM bone marrow, G-CSF granulocyte-colony-stimulating factor, RBC red blood cells

To overcome technical barriers, a few methods to improve the functional human biological systems in mice is to inject humanized mice with recombinant proteins (Huntington et al. 2009; Van Lent et al. 2009), hydrodynamically inject DNA plasmids (Chen et al. 2009), induce lentivirus expression of cytokines (Van Lent et al. 2009), or introduce knock-in gene replacement as so to increase the repertoire of cytokines to support human cells (Billerbeck et al. 2011; Lim et al. 2017; Nicolini et al. 2004; Rongvaux et al. 2011). An example of a technique that is effective does not require complex procedures and can be readily applied in any laboratory is the injection of plasmid DNA (IL-15 and Fms-like tyrosine kinase 3/fetal liver kinase-2 (FLT3/FLK2) ligand) via hydrodynamic tail-vein injection (Chen et al. 2009). Upon application of this method, the expression levels of human cytokines were present for 2–3 weeks, while the levels of functional NK cells remained high for more than a month (Chen et al. 2009). Unlike mice induced to constitutively express cytokines which may activate cells and skew them toward unideal lineages, hydrodynamic injection enables researchers to control the exact timing of cytokine induction, allowing flexible manipulation of the model. On top of this, cytokine-stimulated NK cells expressed activation and inhibitory receptors; attacked in vitro target cells, and responded well to viral infections within an in vivo setting (Chen et al. 2009).

Another method which requires more time and resources to create but eliminates the need for cytokine plasmid injection is the use of transgenic mice with knock-in genes, encoding for cytokines. Four examples of these enhanced immunodeficient mice are, first, NOD.Cg-Prkdcscid Il2rgtm1SugTg (SV40/HTLV-IL3, CSF2) 10-7Jic/JicTac (huNOG-EXL mouse), this strain of super immunodeficient mouse has a high rate of human cell engraftment and expresses both granulocyte/macrophage colony-stimulating factor (GM-CSF) and human IL-3 cytokines, controlled by SV40 promoter, which induces myeloid reconstitution and differentiation.

Second, NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg (CMV-IL3, CSF2, KITLG) 1Eav/MloySzJ (NSG-SGM3 mouse) are knock-in mice expressing IL-3, GM-CSF and stem cell factor (SCF) under the control of human-specific cytomegalovirus (CMV) (Billerbeck et al. 2011; Yao et al. 2016). Even though this combination of genes supports human HSC engraftment, formation of myeloid leukocytes, and reduces B-lymphopoiesis post-BM transplantation this model lacks an improved red blood cell (RBC) reconstitution and the presence of SCF may destructively affect human stem cell compartments by supporting the growth and competitive repopulation of mouse cells (Billerbeck et al. 2011; Yao et al. 2016).

Third, C;129S4-Rag2tm1.1Flv Csf1tm1(CSF1)Flv Csf2/Il3tm1.1(CSF2,IL3)Flv Thpotm1.1(TPO)Flv Il2rgtm1.1FlvTg (SIRPα) 1Flv/J (MISTRG mouse) was designed to support a greater level of human cell reconstitution, particularly in the myeloid compartment by transgenically inducing human GM-CSF, IL-3, macrophage colony-stimulating factor (M-CSF), thrombopoietin (TPO), and signal-regulatory protein alpha (SIRPα). SIRPα produces anti-phagocytic signals upon interaction with human CD47 cells which inhibits murine macrophages from phagocytosing human cells (Rongvaux et al. 2014). However, due to poor erythropoiesis of both mouse and human cells especially post-irradiation preconditioning, MISTRG mice developed severe anemia resulting in its short lifespan and was eventually discontinued commercially (Rongvaux et al. 2014).

Fourth, NOD.Cg-KitW41J Prkdcscid Il2rgtm1Wjl/WaskJ (NSGW41) was created to overcome a lack of erythro-megakaryopoiesis in humanized mouse models. Without the need for irradiation, this KIT-deficient mouse demonstrated improved erythropoiesis and platelet formation as compared to other models of mice (Cosgun et al. 2014; Rahmig et al. 2016). After reconstitution, significant numbers of mature thrombocytes were present in the peripheral blood while human erythroblasts were seen in the BM. In addition, the morphology, composition, and enucleation ability of de novo generated human erythroblasts were similar with those in the human BM (Rahmig et al. 2016). However, as this model is relatively new, more studies are needed to further characterise the advances and limitations of this platform. Details of immunodeficient mouse models are listed in Tables 1 and 2. As existing models are far from perfect, it is important to work on components that enhance cell–cell interactions, support differentiation, and induce maturation of human cells, particularly that of myeloid and B cell compartments to create a model that faithfully recapitulates the human immune system.

Models of Human Diseases Established on Humanized Mice

The introduction of humanized mice provides immeasurable opportunities to advance medical research. These increasingly important pre-clinical models are not only easy to handle due to their small sizes, but they also have short reproductive cycles, an exceptional ability to produce a large number of young and are relatively affordable to maintain in animal facilities as they do not require highly specialised infrastructures that are used by NHPs (Fischer and Austad 2011). In addition, humanized mice allow human-specific pathogens to infect and replicate within them and are able to develop functional human-specific immune responses to an array of diseases.

Many mechanisms underlying diseases are not completely dissected; therefore, utilization of humanized mice allows researchers to understand important factors that facilitate the development of medical issues including infectious disease, cancer, autoimmunity, and GvHD. Currently, a mouse model that completely mimics every single human disease does not exist; therefore, research aims such as the consideration of specific parameters to be analyzed including genotype, phenotype of the model, and scientific budget must be thought through carefully to select a suitable platform.

Infectious Disease

Since the invention of humanized mice, multitudinous attempts have been made to recapitulate infectious diseases within these mice. A particular human-specific infectious pathogen that has been successfully studied on humanized mice is a retrovirus known as human immunodeficiency virus (HIV) (Araínga et al. 2016; Berges and Rowan 2011; Choudhary et al. 2009; Duyne et al. 2011; Li et al. 2014). Before humanized mice were introduced, the only non-human animal model available for dissecting HIV pathogenesis was the chimpanzee (Vanden Haesevelde et al. 1996). Because of cellular and molecular differences between HIV pathogenesis in humans and chimpanzees, restricted tropism of HIV and high-expense of using NHPs, the small, cost-effective, and widely available humanized mice were used in place of the NHPs (Denton and Garcia 2011; Hatziioannou and Evans 2012; Miller et al. 2000).

Humanized mice infected with HIV recapitulated the disease’s progression, latency and virology, permitted long-term immunological studies and helped identify crucial factors such as viral infectivity factor, viral protein u, and negative factor which are essential for in vivo HIV replication (Yamada et al. 2015).

Of all the models (Hu-PBL-scid, Hu-SRC-scid and BLT) (Choudhary et al. 2012; Dash et al. 2011; Gorantla et al. 2010; Ince et al. 2010; Long and Stoddart 2012; Sato et al. 2010; Zhang et al. 2011) used to characterise HIV, BLT mice (Carter et al. 2011; Denton et al. 2012; Marsden et al. 2012) had the most accurate representation of the human mucosal system (Brainard et al. 2009; Denton et al. 2010; Sun et al. 2007), allowing the study of vaginal and rectal transmission and prevention of HIV by enabling evaluations of many prophylactic therapeutics (Balazs et al. 2011), anti-HIV antibodies (Choudhary et al. 2009; Joseph et al. 2010), and cellular therapeutic inventions for inhibiting or eliminating HIV (Holt et al. 2010; Kumar et al. 2008; Neff et al. 2011; Shimizu et al. 2010).

Humanized mouse model with a fully functional human immune system has also been infected with Dengue virus (DENV) (Frias-Staheli et al. 2014; Kuruvilla et al. 2007; Sridharan et al. 2013; Subramanya et al. 2010). These mice demonstrated fever, rash, viremia, erythema, thrombocytopenia, and production of anti-DENV IgM, IgG and a range of cytokines as observed in patients (Mota and Rico-Hesse 2009, 2011). Another human-specific infectious pathogen studied on humanized mice, Plasmodium falciparum, is a causative agent of malaria (Amaladoss et al. 2015; Carlton et al. 2008; Chen et al. 2014; Good et al. 2015; Jiménez-Díaz et al. 2009; Soulard et al. 2015; Vaughan et al. 2012). For years, our understanding of malaria had been impeded by the lack of human-specific small animal models which can be infected by highly host-specific human Plasmodium species (Amaladoss et al. 2015; Chen et al. 2014; Pain et al. 2008) to recapitulate both erythrocytic and immunological disease pathogenesis observed in patients. Due to this, most in vivo experimental studies of malaria were conducted in rodents with mouse or rat-specific Plasmodium strains (Goodman et al. 2013). Differences in invasion and disease pathology between human and rodent parasite species hindered the translation of findings and evaluation of new therapeutics from rodents to humans (Amaladoss et al. 2015; Chen et al. 2014). This challenge has been tackled by incorporating RBC supplemented, immune cell-optimised (enhanced by hydrodynamic expression of human cytokines, IL-15, and FLT3/FLK2 ligand) humanized mice that supports multiple cycles of P. falciparum infection (Amaladoss et al. 2015; Chen et al. 2014).

Utilizing this model, research teams were able to identify the importance of human NK cells, DCs, and B cells in the control of parasitemia. Notably, how NK cells preferentially interacts with infected RBCs (iRBCs), resulting in the activation of NK cells, release of interferon (IFN)-γ, perforin, and granzyme to lyse and eliminate iRBCs in a contact-dependent manner and the importance of adhesion molecule lymphocyte-associated antigen-1 and DNAX accessory molecule-1 which are required for NK cell interaction and clearance of iRBCs (Amaladoss et al. 2015; Chen et al. 2014). Besides facilitating the understanding of human immune responses to Malaria infection, the use of humanized mice also assists in evaluation of new therapeutics and vaccines (Good et al. 2015; Tsuji et al. 1995).

In addition to the human immune system, recent progress has been made to introduce humanization of the liver in humanized mice to support the study of hepatotropic pathogens such as hepatitis B virus and hepatitis C virus (HCV) (Bility et al. 2012; Keng et al. 2015; Strick-Marchand et al. 2015; Tan-Garcia et al. 2017; Washburn et al. 2011). It has been shown that these new humanized mice could be infected with human strains of hepatitis viruses and exhibit leukocyte infiltrations, liver inflammation, fibrosis, cirrhosis, and elevated cytokines similar to HCV-infected patients (Bility et al. 2014; Keng et al. 2015; Tan-Garcia et al. 2017; Washburn et al. 2011). Mouse models with human liver cells and matched human immune system provides an important platform for understanding disease pathogenesis of hepatitis viruses through human-specific cytokines, chemokines and immune cell regulations involved, potentially translating this knowledge into creation of anti-fibrotic and immune-modulatory therapeutics (Bae et al. 2015; Keng et al. 2015).

Other examples of infectious pathogens studied on humanized mice include, Mycobacterium tuberculosis (Calderon et al. 2013; Nusbaum et al. 2016), influenza (Yu et al. 2008; Zheng et al. 2015), Borrelia hermsii (Vuyyuru et al. 2011), human CMV (Daenthanasanmak et al. 2015; Smith et al. 2010), Ebola virus (Bird et al. 2016; Lüdtke et al. 2015), Epstein-Barr virus (Cocco et al. 2008; Sato et al. 2011; Yajima et al. 2008) and Kaposi’s sarcoma-associated herpesvirus (Boss et al. 2011; Chang et al. 2009; Wang et al. 2014). Further details on infectious pathogens that have been studied using humanized mice as a platform are detailed in Table 4.

Table 4

Infectious diseases modelled in humanized mice

Infectious disease

Model

Main findings

References

Borrelia hermsii

Newborn NSG engrafted with human CD34+ UBC cells within 48 h of birth and intravenously or intraperitoneally infected with B. hermsii

Similar to clinical scenarios, infection of humanized mice with B. hermsii resulted in recurrent episodes of bacteremia which was resolved with B. hermsii specific IgM production. Anti-B. hermsii responses were diminished and persistent bacteremia recurred upon administration of anti-human CD20 antibody

Vuyyuru et al. (2011)

DENV

NOD/scid engrafted with human fetal thymus and liver tissue under the kidney capsule and intravenously injected with CD34+ human FL cells to create huBLT mice. Mice were intravenously infected with DENV-2

Intravenous inoculation of DENV-2 resulted in sustained viremia and infection of leukocytes in lymphoid and non-lymphoid organs. Serum cytokine levels and DENV-2-neutralising human IgM antibodies were detected in infected mice. In re-stimulation with DENV-infected DCs, in vivo primed T cells were activated and had effector functions

Frias-Staheli et al. (2014)

Ebola virus

NSG-A2 intravenously (retro-orbital) injected with human CD34+ UBC from HLA-A2 donors and intraperitoneally infected with Ebola virus

Similar to clinical scenarios, mice showed signs of viremia, cell damage, liver steatosis, and hemorrhage

Lüdtke et al. (2015)

EBV

NOG mice intravenously injected with human CD34+ UBC and EBV

B cell lymphoproliferative disorder was observed with high dose of EBV. Low dose of EBV resulted in asymptomatic persistent infection, increased levels of CD8+ T in the peripheral blood, EBV-specific T cell responses and IgM specific to EBV-encoded protein BFRF3

Yajima et al. (2008)

HBV

NSG-A2 mice were intrahepatically injected with autologous CD34+ HSC and hepatic progenitor cells to create A2/NSG-hu HSC/Hep mice. These mice were intravenously infected with clinical isolates of HBV

Mice were able to demonstrate persistent infection for up to 4 months after HBV inoculation. Similar to clinical scenarios, chronic liver inflammation, liver fibrosis and immune responses were observed in infected mice. Neutralising antibody (anti-HBsAg scFv) was able inhibit liver disease

Bility et al. (2014)

HCV

Newborn NSG were intrahepatically injected with human CD34+ FL cells within 72 h of birth and intravenously infected with HCV

Humanized mice were able to support HCV infection and demonstrated clinical symptoms and immune responses (innate and adaptive) commonly observed in HCV-infected patients

Keng et al. (2015)

hAdV

HLA-A2 mice were engrafted with autologous human CD34+ HSPCs from UCB via intra-orbital injection and intravenously infected with hAdV

Humanized mice recapitulated the pathology of acute and persistent hAdV infection. In acute infection, high mortality, weight loss, liver pathology and expression of viral protein within organs were observed. Chronic infection was asymptomatic and resulted in the development of hAdV-specific adaptive immunity and expression of early viral genes within the BM

Rodríguez et al. (2017)

hCMV

NRG mice engrafted with CD34+ human cells isolated from adult PBMCs and UBC and infected with hCMV

When a tricistronic integrase-defective lentiviral vector (co-expressing GM-CSF, IFN-α, and hCMV pp65 antigen) which induced self-differentiation of monocytes in PBMCs and UCB into DCs with pp65 (“SmyleDCpp65”) was administered, humanized mice infected with hCMV demonstrated remodeling of LNs, upregulation of thymopoiesis in CD4+ and CD8+ T cell precursors, polyclonal effector memory CD8+ T cells expansion in blood, spleen, and BM, PP65-specific CTL, and IgG responses

Daenthanasanmak et al. (2015)

HIV

Newborn NSG intrahepatically injected with CD34+ human FL cells and infected with HIV-1ADA via intraperitoneal injection

Cell distribution and HIV viral life cycle were dependent on tissue compartment and time of infection. HIV-1 in cells was found as forms of integrated DNA and multi- and un-spliced RNA

Araínga et al. (2016)

HTLV1

NOG mice engrafted with human CD133+ UBC cells by IBMI) to create IBMI-huNOG mice which were intraperitoneally infected with HTLV-1

Infected mice recapitulated symptoms of adult T-cell leukemia and HTLV-1-specific adaptive immune responses including, elevation of CD4+ T cells, and signs of atypical lymphocytes with lobulated nuclei

Tezuka et al. (2014)

Influenza

Rag2−/−γc−/− mice intraperitoneally injected with human PBMCs and Intranasally infected with Influenza

Intraperitoneal injection of pamidronate induced Vδ2-T cells to secrete IFN-γ and kill virus infected host cells which helped to control viral replication and suppressed inflammation in lungs of H7N9-infected mice, reducing their morbidity and mortality

Zheng et al. (2015)

KSHV

NSG mice engrafted with human fetal thymus and liver tissue under the kidney capsule and intravenously injected with CD34+ human FL cells to create huBLT mice. Mice were infected with KSHV via the oral mucosa

Mice were infected with KSHV via the oral mucosa and established a robust infection by targeting human macrophages and B cells

Wang et al. (2014)

Leishmania major

Newborn NSG intrahepatically injected with human CD34+ UBC cells and infected with Leishmania major via subcutaneous footpad injection

At the site of injection, human macrophages were infected with Leishmania parasites and Leishmania-specific human T cell responses were detected. Miltefosine reduced parasitic load and induced side-effects as observed in clinical scenarios

Wege et al. (2012)

Malaria

Newborn NSG intracardially injected with human CD34+ UBC cell and intravenously infected with malaria

NSG mice were supplemented human erythropoietin and IL-3 via hydrodynamic tail-vein injection. Human RBCs generated de novo were infected with P. falciparum and it was observed that different strains of parasites varied in their infection rates

Amaladoss et al. (2015)

NiV

NSG mice engrafted with human lung tissue and intragraft injected with NiV

Human fetal lung xenografts were able to form human adult lung structures. NiV replicated to high titers and infected human lung tissues resulting in the production of cytokines and chemokines including IL-6, G-CSF, and GM-CSF which commonly causes acute lung injury

Valbuena et al. (2014)

Mycobacterium tuberculosis

NSG mice engrafted with human fetal thymus and liver tissue under the kidney capsule and intravenously injected with CD34+ FL cells to create huBLT mice. These mice were intranasally infected with tdTomato M. tuberculosis H37Rv

Mice infected with M. tuberculosis demonstrated progressive bacterial infection within the lung which disseminated to the spleen and liver. Pathological analysis of the infected lung displayed obstruction of the bronchial, granulomatous lesions, caseous necrosis and crystallised cholesterol deposits. Human T cells were detected at sites of inflammation and bacterial growth, within the lung, liver, and spleen

Calderon et al. (2013)

VZV

NOD/scid mice engrafted with human fetal thymus and liver tissue under the kidney capsule or subcutaneously implanted with fetal skin. MRC-5 cells infected with wild-type VZV/Oka strain was injected into the implants

Varicella-zoster viral proteins were expressed in CD4+ and CD8+ T cells which have a capacity to cause viremia. Similar to clinical scenarios, skin implants infected with VZV showed lesions of varicella

Moffat et al. (1995)

DENV Dengue virus, EBV Epstein–Barr virus, HBV hepatitis B virus, HCV hepatitis C virus, hAdV human adenovirus, hCMV human cytomegalovirus, HIV human immunodeficiency virus, HTLV1 human T-lymphotropic virus 1, KSHV Kaposi’s sarcoma-associated herpesvirus, NiV Nipah virus, VZV Varicella-zoster virus, BLT bone marrow/liver/thymus, HSC hematopoietic stem cells, FL fetal liver, PBMCs peripheral blood mononuclear cells, UCB umbilical cord blood, BM bone marrow, GM-CSF macrophage granulocyte-colony-stimulating factor, IBMI intra-BM injection, HSPCs hematopoietic stem and progenitor cells, DCs dendritic cell, IFN interferon, LNs lymph nodes, scFv single-chain variable fragment, CTL cytotoxic T lymphocyte, RBC red blood cell, tdTomato Tandem dimer Tomato

Cancer

Immunodeficient mice that lack innate and adaptive immune cell compartments enable successful engraftment of many human tumors including tumor cell lines and primary solid and hematological tumors. Currently, there are three ways to study tumor growth and cancer immunology in humanized mice. First, tumor cell lines can be engrafted into humanized mice reconstituted with HSCs or PBMCs (Ito et al. 2009; Tsoneva et al. 2017; Wege et al. 2014). Breast cancer was modelled in mice by concurrently transplanting CD34+ HSCs and tumor cells into newborn mice or engrafting both PBMCs and tumor cells into BRG mice (Wege et al. 2014). In these models, human immune cells were able to traffic and infiltrate the microenvironment, enabling human tumor-immune system interactions to be studied (Wege et al. 2014). To more closely recapitulate human immune responses to tumor cell lines, MISTRG mice engrafted with CD34+ human FL cells were subcutaneously transplanted with a melanoma cell line, Me290 (Rongvaux et al. 2014). Similar to clinical scenarios, it was observed that myeloid cells infiltrated the tumor, numerous cells within the tumor expressed CD14 and CD163 which are commonly associated as macrophage markers, and CD163+ cells were most likely M2-like macrophages as they were HLA-DRlow and CD206high. It was hypothesised that tumor growth may have been mediated by M2-like macrophages that can induce cytokine production or release enzymes to promote vascularisation and immune suppression. Therefore, these mice were treated with human-vascular endothelial growth factor (VEGF) inhibitor, Avastin®. Humanized mice engrafted with Me290 responded to treatment by inhibiting tumor growth, suggesting that myeloid cells may support tumor growth via VEGF activity (Rongvaux et al. 2014).

Second, immunodeficient mice can be engrafted with patient-derived xenografts (PDX) (Bankert et al. 2011; Her et al. 2017; Simpson-Abelson et al. 2008). Engraftment of patient-derived acute myeloid leukemia (AML) cells into newborn NSG resulted in high levels of human cell engraftment in the peripheral blood, spleen and BM of recipient mice (Her et al. 2017). Similar to observations in the clinics, these mice also had enlarged spleens and infiltration of AML cells into multiple organs. Even though AML remained unaltered during serial transplantation, many studies with engrafted PDXs into immunodeficient mice have demonstrated that heterogeneity of parental tumor was often only maintained in primary engraftment (Cassidy et al. 2015). Over time and tumor passage, human stromal was frequently compromised by infiltration and replacement with mouse-derived cells (Cassidy et al. 2015; Maykel et al. 2014). This model is ideal for understanding stroma–tumor interactions, which is integral for tumor growth and an important target for cancer therapy.

Third, for a comprehensive study of interactions between human immune cells and tumor in vivo, immunodeficient mice should be engrafted with PDX and human immune cells (Pan et al. 2017; Roth and Harui 2015). This humanized PDX model would not only have a complete tumor microenvironment but also an ability to display heterogeneity lost in tumors (Pan et al. 2017). However, a drawback of this model is the scarcity of autologous HSCs which affects the capacity to generate cohorts for research. To overcome this challenge, HSCs isolated from UBC, FL or G-CSF mobilised PBMCs can be expanded either by transduction with tat-MYC and tat-Bcl2 fusion proteins or cultured with a validated cocktail of growth factors to induce in vitro proliferation of HSCs (Bird et al. 2014; Yong et al. 2016). An example of this model is XactMice which are engrafted with in vitro expanded HSCs and autologous PDX samples from head and neck squamous cell carcinoma patients (Morton et al. 2016). Even though these mice had low levels of humanization in their peripheral blood, they demonstrated an increase in lymphatic vessels and the presence of CD45+CD151+ cells, suggesting that these mice were able to recapitulate immune and stromal cell compartments of the tumor microenvironment (Morton et al. 2016).

While the current immunodeficient mouse strains are able to support the engraftment of most tumor cell lines, not all primary tumors for example prostate cancer can be easily engrafted (Roth and Harui 2015). Novel humanized oncological models are being innovated to address important questions on tumor-immune system interactions, mechanisms of tumor escape, therapeutic potential of immune modulation, as well as refining therapeutic solutions such as chemotherapy, NK cell therapy, checkpoint inhibitors and cytokine therapy. Tumor cell lines, and solid and hematological cancers tested on humanized mice are listed in Table 5.

Table 5

Cancer modelled in humanized mice

Cancer

Model

Main findings

References

Bladder

NSG mice were injected with CD34+ hematopoietic progenitor cells and subcutaneously engrafted with patient-derived bladder cancer cells

Major human immune cell subsets were reconstituted in humanized mice, no xenograft-versus-host disease was observed and PDX retained morphological and genetic fidelity of parental patient cancer

Pan et al. (2017)

Breast

NSG were intrahepatically engrafted with human breast carcinoma cell line (SK-BR-3)

Mice were engrafted with functional human immune system and human breast cancer cells. MHC-mismatched tumor cells resulted in activated immune cells, but no clinical signs of rejection were observed

Wege et al. (2014)

Cervical

Human cervical carcinoma cell line (C33a) was subcutaneously engrafted into scid mice

Herpes simplex virus type I-based oncolytic treatment in combination with radiation therapy may be an effective treatment for cervical cancer

Blank et al. (2002)

Colorectal

Rag2−/−γc−/− mice were injected with human PBMCs and subcutaneously engrafted on the flank with colorectal carcinoma cell line (HT-29)

Co-administration of Urelumab and Nivolumab slowed down tumor growth by elevating activated human T lymphocytes which produced IFN-γ and decreased levels of human regulatory T cells in tumor xenografts

Sanmamed et al. (2015)

Gastric

Patient-derived xenografts of gastric cancer were subcutaneously engrafted into the right hind flank of scid and nude mice

Mice engrafted with patient-derived gastric cancers demonstrated identical histological and genetic diversities which corresponded to parental patient tumors

Zhang et al. (2015)

HNSCC

NSG mice were injected with expanded HSPCs and engrafted with patient-derived HNSCC

Human immune and stromal cells produced in XactMice mimics patient’s tumor microenvironment. This model was able to reverse genetic drift of tumors that usually occur after serial transplantation in non-humanized mice

Morton et al. (2016)

Kidney

NSG mice were engrafted with human RCC cell line (SKRC-59 cells) in the left subrenal capsule of their kidney

Human anti-CAIX mAbs inhibit RCC growth by halting migration and triggering immune-mediated killing of RCC. Improvements to anti-CAIX mAbs demonstrated enhanced antibody-dependent cell-mediated cytotoxicity against RCC

Chang et al. (2015)

Leukemia

Newborn NSG were intravenously engrafted with patient-derived AML cells

High levels of AML engraftment were observed in the peripheral blood, spleen and BM of recipient mice. Similar to clinical scenarios, mice had enlarged spleen and infiltration of AML cells into multiple organs. Serial transplantation did not alter AML cells

Her et al. (2017)

Lung

NSG and C.B-17-scid subcutaneously engrafted with patient-derived xenograft at a position caudal to the xiphoid process

NSG mice were successfully engrafted with patient-derived primary lung tumors. Mice retained parental tumor architecture such as tumor-associated leukocytes, stromal fibroblasts, and had limited xenograft-versus-host disease. Tumor-associated T cells migrated from the microenvironment of xenografts toward the lung, liver, and spleen of mice

Simpson-Abelson et al. (2008)

Lymphoma

NOG mice were subcutaneously engrafted with human PBMCs and injected with Hodgkin lymphoma cell line (L-428) or cutaneous T-cell lymphoma cell line (HH)

Anti-CCR4 mAb KM2760 demonstrated anti-tumor activity in humanized mouse models of lymphoma. Upon treatment of KM2760, tumor-infiltrating CD56+ NK cells were increased and T-regulatory cells were decreased

Ito et al. (2009)

Melanoma

Newborn NSG were intrahepatically injected with CD34+ UBC and injected with human melanoma cell lines (1935-MEL and 888-MEL)

Mice were successfully engrafted with a functional human immune system. Oncolytic vaccinia virus therapy, particularly CTLA4 scAb increased CD56+ NK cells and decreased virus titers

Tsoneva et al. (2017)

Myeloma

NOG mice were intravenously engrafted with human myeloma cell lines (U266)

U266 myeloma cells homed to the BM and resulted in paralysis of NOG mice

Miyakawa et al. (2004)

Ovarian

NSG mice were intraperitoneally engrafted with patient-derived xenografts of primary and metastatic ovarian solid tumor tissue and ovarian ascites fluid

Similar to clinical patients, tumors engrafted in these mice established in the omentum, ovaries, liver, spleen, uterus, and pancreas

Bankert et al. (2011)

Pancreatic

NSG mice were engrafted with patient-derived pancreatic cancer tumors by subcutaneous, intravenous or intra-pancreatic injections

Activated allogenic and autologous NK cells were able to selectively kill cancer stem cells in NSG mice engrafted with pancreatic cancer

Ames et al. (2015)

Prostate

NSG mice were injected with PBMCs with subsets of CD4+, CD8+ and autologous DCs and subcutaneously injected with human prostate cancer cells (PC3) into the right flank

Tumor-infiltrating lymphocytes in NSG mice with a functional human immune system and prostate cancer cells were similar to clinical scenarios

Roth and Harui (2015)

HNSCC head and neck squamous cell carcinoma, RCC renal cell carcinoma, AML acute myeloid leukemia, PDX patient-derived xenografts, mAbs monoclonal antibodies, scAb single-chain antibody

Autoimmunity

Disparities in the immune system between mice and men restrict the use of mouse models which develops spontaneous autoimmunity (Covassin et al. 2013). To overcome this challenge, Gunawan et al. (2017) engrafted PBMCs from systemic lupus erythematosus (SLE) patients to create a human-specific disease-based immune system which demonstrated that human T and B cells were present in the peripheral blood and spleen of humanized mice and were important to lupus development. Similar to patients, when these mice were treated with dexamethasone, spleen weight, and proteinuria decreased. Mice with a human immune system xenografted with patient samples allow a spectrum of disorders such as SLE (Andrade et al. 2011; Gunawan et al. 2017) and type I diabetes (Shultz et al. 2007; Unger et al. 2012; Viehmann Milam et al. 2014) to be evaluated for the identification of screening markers, retrieval of antigen-specific autoantibodies, and drug tests. Autoimmune diseases that have been studied using humanized mice as a platform are listed in Table 6.

Table 6

Autoimmune diseases modelled in humanized mice

Autoimmunity

Models

Main findings

References

Multiple sclerosis

NSG mice engrafted with PBMCs and injected with myelin antigens in Freund’s adjuvant and antigen-pulsed autologous DCs

Mice demonstrated subclinical CNS inflammation. Human T cells (CD4+ and CD8+) were specific to the soluble domain of myelin oligodendrocyte glycoprotein and produced proinflammatory cytokines

Zayoud et al. (2013)

SLE

NSG mice engrafted with FL HSCs and injected with pristane

Humanized mice recapitulated key clinical and immunological features of SLE including production of human anti-nuclear autoantibodies, lupus nephritis, pulmonary serositis, decreased human lymphocytes in peripheral blood, hyperactivated B and T cells and increased proinflammatory cytokines

Gunawan et al. (2017)

SjS

NSG mice engrafted with PBMCs from patients with SjS

Mice engrafted with PBMCs from SjS patients had elevated levels of cytokines, particularly IFN-γ and IL-10. Histological analysis showed signs of inflammation within the lacrimal and salivary glands of mice engrafted with SjS. These infiltrates were mostly CD4+ and a small population of CD8+ T cells and B cells

Young et al. (2015)

Type I diabetes

NSG-Abo DR4 engrafted with CD4+ T cells pulsed with autoantigen-derived peptides

Mice injected with autoantigen-reactive CD4+ T cells lines from diabetic donors demonstrated human T cells infiltration into mouse islets, insulitis, and increased levels of demethylated β-cell–derived DNA in the bloodstream and reduced levels of insulin staining

Viehmann Milam et al. (2014)

SLE Systemic lupus erythematosus, SjS Sjogren’s syndrome, CNS central nervous system

Graft-versus-host Disease

The occurrence of GvHD is a life-threatening complication that may develop following transplantations (Hu et al. 2011; Hu and Yang 2012). Even though GvHD has been intensively analyzed in non-humanized animal models, many human-specific mechanisms and treatments cannot be tested due to incongruence between humans and mice. Humanized mice are excellent substitutes to investigate exact human immune responses of GvHD and its related therapeutics (Ali et al. 2012; King et al. 2008; Kirkiles-Smith et al. 2009; Tobin et al. 2013; Wang et al. 2011; Zhao et al. 2015). An example of a humanized mouse model applied in GvHD studies is the engraftment of human PBMCs into immunodeficient mice (Ali et al. 2012). Post-transplantation, these mice demonstrated human lymphocytes infiltration into peripheral blood, spleen, lymph nodes, and BM of the mice, had enhanced tissue homing cells with a T-effector memory (TEM) phenotype and high levels of cutaneous lymphocyte antigen, recapitulating the exact pathogenesis of GvHD as observed in patients (Ali et al. 2012; Wang et al. 2011). Utilizing humanized mice to understand human-specific mechanisms of rejection provides a strong pre-clinical platform for the design of novel immunotherapies (Fogal et al. 2011; Onoe et al. 2011; Tobin et al. 2013), especially those targeting TEM cell driven GvHD (Ali et al. 2012). Transplant rejection studies that have been conducted on humanized mice are listed in Table 7.

Table 7

GvHD modelled in humanized mice

GvHD

Models

Main findings

References

Cardiac tissue and skin

NSG mice were engrafted with human skin and artery tissue and injected with enriched human CD34+ HSC isolated from peripheral blood of G-colony stimulated factor pre-treated adults or PBMCs autologous to CD34+ donors either separately or together

Without T cells, CD14+CD68+ macrophages infiltrate allogeneic human skin but caused minimal injury and thrombosis. However, with the adoptive transfer of T cells autologous to HSC, CD14+CD68+ macrophages infiltrated allogeneic arterial interposition grafts, induced intimal expansion and calcification

Kirkiles-Smith et al. (2009)

hiPSCs

NSG mice engrafted with human fetal thymus and liver tissue under the kidney capsule and intravenously injected with autologous CD34+ human FL cells to create huBLT mice

Signs suggesting immune rejection of hiPSCs including formation of teratoma, infiltration of antigen-specific T cells and tissue necrosis were observed in these mice engrafted with autologous integration-free hiPSCs. In this study, autologous hiPSC-derived smooth muscle cells were highly immunogenic, while autologous hiPSC-derived retinal pigment epithelial cells were immune tolerated

Zhao et al. (2015)

Islet

NSG injected with human PBMCs and engrafted with human islets

Mice demonstrated low intra- and inter-donor variability of PBMCs engraftment. When treated with streptozotocin, mice were hyperglycemic but returned to normoglycemia when transplanted with islet cells. Upon injection of HLA-mismatched human PBMCs, mice showed signs of hyperglycemia, loss of human C-peptide, and rejection of human islet grafts

King et al. (2008)

PBMCs

NSG mice injected with human PBMCs alone or incubated with MSCs or stromal cells

Effectiveness of MSC therapy was dependent on the time of administration. Mice demonstrated signs of reduced liver and gut pathology and increased survival. MSC therapy did not result in donor T cell anergy and regulatory T cells did not induce the apoptosis of PBMCs; instead, it was associated with direct inhibition of donor CD4+ T cell proliferation and reduction of human TNF-α within the serum

Tobin et al. (2013)

GvHD graft-versus-host disease, hiPSCs human induced pluripotent stem cells, PBMCs peripheral blood mononuclear cells, MSCs mesenchymal stem cells, HSC hematopoietic stem cell, TNF tumor necrosis factor

Human-Specific Drug Tests on Humanized Mouse Models

Non-human animal models are commonly used to test an array of human-specific therapeutics during pre-clinical trials. Due to a lack of human specificity, it is common for pre-clinical trials to inadequately identify exact pharmacokinetics, pharmacodynamics, and side-effects of therapeutics, which may result in debilitating and life-threatening situations when tested on humans (Horvath et al. 2012; Rehman et al. 2011; Xu et al. 2014). To improve from unsuccessful clinical trials, it is important to use validated and cost-effective animal models with high human specificity such as humanized mouse models to expand the traditional armamentarium of therapeutics for treatment of patients with complicated and progressive conditions.

Therapeutics successfully tested in mice with a functional human immune system includes an antiviral drug, peginterferon alpha-2a (Peg-IFNα2a) which demonstrated signs of HCV inhibition such as decreased human IFN-γ production, level of serum alanine aminotransferase, copies of HCV ribonucleic acid (RNA), and absence of leukocyte infiltration or fibrosis in the liver (Keng et al. 2015). Similar to clinical scenarios, humanized mice administered with Ipilimumab developed autoimmune disease with signs of weight loss, anti-nuclear antibodies, and adrenalitis. In addition, a biologic highly specific for human CD28, theralizumab, was tested in humanized mice engrafted with PBMCs (Weißmüller et al. 2016). These mice demonstrated severe reduction in CD45+ human cells, rapid drop of body temperature, elevated levels of cytokines, and succumbed to treatment within 6 h after antibody administration, recapitulating adverse effects observed in clinical scenarios (Weißmüller et al. 2016).

Considering the strengths, limitations, and potential developments of humanized mice, the current data indicate that these models are beneficial tools for researchers to investigate short and long-term studies of in vivo therapeutic interactions and toxicities to mitigate risks and ensure the safety of healthy volunteers and patients exposed to candidate agents during clinical trials. Therapeutics that has been tested on humanized mice is listed in Table 8.

Table 8

Therapeutics tested on humanized mice

Therapeutic

Alternative names

Model

Main findings

References

Alemtuzumab

Campath®, Campath-1H, MabCampath and Lemtrada

NSG mice intravenously injected with human PBMCs

Similar to clinical scenarios, Alemtuzumab induced severe temperature reduction in mice and bound to CD3 and CD52 but did not induce activation of markers CD25 and CD69

Brady et al. (2014)

ATG

Thymoglobulin®

NSG mice injected with human PBMCs

Mice that were given 150 µg of ATG intravenously became sick and were sacrificed within 1 h after treatment. Optimal dose of ATG in this study was 30 µg, where mice demonstrated mild clinical signs of drug treatment but recovered within 5 h

Brady et al. (2014)

Eltrombopag

Promacta®, Revolade

NOD/scid mice intravenously injected with human CD34+ UCB cells

Eltrombopag enhanced expansion and promoted multilineage hematopoiesis of HSPCs

Sun et al. (2012)

Ipilimumab

Yervoy®

Newborn NSG were intrahepatically injected with human CD34+ FL/UCB cells within 24 h of birth

Ipilimumab accelerated rejection of skin graft on humanized mice

Waldron-Lynch et al. (2012)

KM2760

NOG mice were engrafted with human PBMCs and injected with Hodgkin lymphoma cell line (L-428) or cutaneous T-cell lymphoma cell line (HH)

Anti-CCR4 mAb could be used to induce anti-tumor activity by removing CCR4-expressing tumors and downregulating regulatory T cells

Ito et al. (2009)

Lamivudine

3TC

C.B-17-scid engrafted with human thymus and liver tissues under the kidney capsule (scid-hu Thy/Liv mouse)

Relative to untreated mice, intraperitoneal injection of 3TC at 30 mg/kg/day had large reductions in viral RNA from a mean of 104.7 to 101.8 copies per 106 cells

Stoddart et al. (2014)

Miltefosine

Impavido

Newborn NSG were engrafted with human CD34+ UBC cells and injected with stationary phase promastigote L. major into the footpad

Parasitic load was reduced and humanized mice demonstrated side-effects similar to clinical scenarios

Wege et al. (2011)

Muromonab-CD3

Orthoclone OKT3

NSG mice intravenously injected with human PBMCs

Administration of Muromonab-CD3, particularly intravenously resulted in cytokine storm and acute clinical symptoms such as piloerection, hypomotility and hypothermia

Brady et al. (2014)

Nivolumab

Opdivo®

RAG2−/−γc−/− mice intravenously injected with human PBMCs

In mice engrafted with human colorectal HT-29 carcinoma cells and allogeneic human PBMCs, co-administration of Nivolumab and Urelumab slowed tumor growth

Sanmamed et al. (2015)

Oseltamivir

Tamiflu®

RAG2−/−γc−/− mice intraperitoneally injected with H7N9

No therapeutic effects were observed when humanized mice were infected H7N9 were treated with Oseltamivir

Zheng et al. (2015)

Pamidronate

Aredia®

RAG2−/−γc−/− mice intraperitoneally injected with H7N9

Pamidronate induced controlled viral replication and suppressed H7N9 injected within humanized mice. Treating mice with Pamidronate 3 days after infection could still ameliorate the disease

Zheng et al. (2015)

Peg-IFNα2a

Pegasys®

Newborn NSG were intrahepatically injected with human CD34+ FL cells within 72 h of birth

HCV copy numbers and serum ALT levels were reduced and no leukocyte infiltrations or fibrosis were observed in HCV-infected humanized mice intramuscularly injected with Peg-IFNα2a

Keng et al. (2015)

PG9

C.B-17-scid engrafted with human thymus and liver tissues under the kidney capsule (scid-hu Thy/Liv mouse)

PG9 provides minimal protective functions in scid-hu Thy/Liv mice challenged with HIVNL4−3. Antibodies can penetrate tissues to prevent infection

Stoddart et al. (2014)

PG16

NSG-BLT mice intravenously injected with human CD34+ FL cells

Single dose of PG16 administered a day before inoculation of HIV was effective in preventing infection

Stoddart et al. (2014)

Regorafenib

Stivarga®

Newborn NSG engrafted with patient primary AML cells

Regorafenib reduced the amount of engrafted human cells within the peripheral blood, extent of myeloid sarcoma and spleen size in mice injected with AML cells

Her et al. (2017)

Sorafenib

Nexavar®

Newborn NSG engrafted with patient primary AML cells

Sorafenib drastically reduced human cells in the peripheral blood, therefore, minimalising the extent of myeloid sarcoma and reducing spleen size in AML mouse model

Her et al. (2017)

Teplizumab

MGA031, hOKT3γ1(Ala-Ala)

Newborn NSG were intrahepatically injected with human CD34+ FL/UCB cells within 24 h of birth

Teplizumab delayed rejection of skin graft on humanized mice

Waldron-Lynch et al. (2012)

Theralizumab

TGN1412, CD28-SuperMAB and TAB08

NRG mice intravenously injected with human PBMCs

Similar to clinical scenarios, humanized mice had a rapid decrease in body temperature, became sick and succumbed to TGN1412, 2–6 h after antibody administration

Weißmüller et al. (2016)

Truvada

(Combination of Tenofovir disoproxil fumarate and Emtricitabine)

C.B-17-scid engrafted with human thymus and liver tissues under the kidney capsule (scid-hu Thy/Liv mouse)

A large dose of Emtricitabine is results in only a small reduction of HIV RNA in HIVJR−CSF-challenged mice

Stoddart et al. (2014)

Urelumab

RAG2−/−γc−/− mice intravenously injected with human PBMCs

Administration of both Urelumab and Nivolumab slowed tumor growth in mice engrafted with HT-29 colorectal carcinoma cells and allogenic human PBMCs

Sanmamed et al. (2015)

ATG anti-thymocyte globulin, HSPCs hematopoetic stem and progenitor cells

Future Directions and Conclusion

To address gaps in humanized mice, scientists working in different biomedical disciplines are attempting a myriad of approaches including boosting human cell reconstitution, reducing graft rejections, supporting critical immune cell subsets, and improving human-specific responses toward pathogens to maximise the potential of humanized mice as a pre-clinical platform. Despite an optimistic outlook of humanized mice, there are considerable obstacles associated with the model that has to be solved as soon as possible. This includes scarce sources of human cells and tissues, particularly obtained from fetal samples due to ethical restrictions. A solution for this limitation is underway as teams around the world perfect induced pluripotent stem cell (iPSC) technology, which enables the use of patient-specific iPSCs allowing a renewable source of autologous cells sans immune rejection (Shi et al. 2017).

In humanized mice, secondary lymphoid structures are either missing or disorganised; this curtails essential humoral responses, resulting in impairments for both class switching and affinity maturation post-immunisation. To overcome this, lymphoid tissue inducer cells should be introduced without affecting IL2rg receptors (Lim et al. 2017). Alternatively, immunodeficient mice can be engrafted with both FL and cells that support FL cell growth from the same clinical donor and supplemented with cytokines (e.g., IL-1β, IL-2, IL-7, and GM-CSF), so that differentiation and maturation of HSCs can take place to improve functional immune cells including macrophages, follicular DC, and T helper cell reconstitution (Chen et al. 2009; Lim et al. 2017; Yong et al. 2016).

An absence of essential human cytokines hinders optimal HSC engraftment, differentiation, and maturation of functional immune cells. To tackle this issue, mouse models can be hydrodynamically boosted with plasmids encoding cytokines (Chen et al. 2009). Despite this improvement, binding of human cytokines may be hindered by residual mouse cytokines or may induce mouse cells to proliferate and displace the engraftment of human cells due to the cross-reactivity between some human and mouse cytokines. Eliminating this problem entirely would require absolute depletion of murine cells or the introduction of high affinity human-specific cytokines and growth factors.

Human cell engraftment is being negatively affected by mouse cells (RBCs and innate immune cells) that were not completely depleted during the construction of immunodeficient mice. To improve this, additional gene knock-outs could be added to current strains of immunodeficient mice to further reduce mouse RBCs, granulocytes and macrophage functions (Hu et al. 2011; Hu and Yang 2012), however, because of the low human erythrocyte engraftment, excessive reduction of mouse RBCs might result in anemic mice which has short lifespans, are weak and not suitable for experiments (Rongvaux et al. 2014). A long-term solution would be to optimise and increase the engraftment rate of human RBCs in humanized mice, so that all traces of mouse RBCs can be removed (Hu and Yang 2012).

Long-termism, critical analysis, and adequate troubleshooting to solve existing problems in humanized mice would undoubtedly provide exciting opportunities for the establishment of new and improved humanized models with increased human immune cell engraftment and enhanced functionality that would greatly benefit the community.

Notes

Acknowledgements

This work was supported by the following grants: National Research Foundation Fellowship Singapore NRF-NRFF2017-03 (Q. Chen.), Eradication of HBV TCR Program: NMRC/TCR/014-NUHS/2015, National Medical Research Council, Singapore (Q. Chen) and A*STAR graduate scholarship from Agency for Science, Technology and Research (A*STAR), Singapore (K.S.M. Yong).

References

  1. Akkina R (2013) New generation humanized mice for virus research: comparative aspects and future prospects. Virology 435:14–28PubMedPubMedCentralCrossRefGoogle Scholar
  2. Ali N, Flutter B, Sanchez Rodriguez R et al (2012) Xenogeneic graft-versus-host-disease in NOD-scid IL-2Rγnull mice display a T-effector memory phenotype. PLoS One 7:e44219PubMedPubMedCentralCrossRefGoogle Scholar
  3. Amaladoss A, Chen Q, Liu M et al (2015) De novo generated human red blood cells in humanized mice support Plasmodium falciparum infection. PLoS One 10:e0129825PubMedPubMedCentralCrossRefGoogle Scholar
  4. Ames E, Canter RJ, Grossenbacher SK et al (2015) NK cells preferentially target tumor cells with a cancer stem cell phenotype. J Immunol 195:4010–4019PubMedPubMedCentralCrossRefGoogle Scholar
  5. Andrade D, Redecha PB, Vukelic M et al (2011) Engraftment of PBMC from SLE and APS donors into BALB-Rag2/IL2Rgc/ mice: a promising model for studying human disease. Arthritis Rheum 63:2764–2773PubMedPubMedCentralCrossRefGoogle Scholar
  6. Araínga M, Su H, Poluektova LY et al (2016) HIV-1 cellular and tissue replication patterns in infected humanized mice. Sci Rep 6:23513PubMedPubMedCentralCrossRefGoogle Scholar
  7. Aryee KE, Shultz LD, Brehm MA (2014) Immunodeficient mouse model for human hematopoietic stem cell engraftment and immune system development. Methods Mol Biol 1185:267–278PubMedPubMedCentralCrossRefGoogle Scholar
  8. Attarwala H (2010) TGN1412: From discovery to disaster. J Young Pharm 2:332–336PubMedPubMedCentralCrossRefGoogle Scholar
  9. Bae KH, Lee F, Xu K et al (2015) Microstructured dextran hydrogels for burst-free sustained release of PEGylated protein drugs. Biomaterials 63:146–157PubMedCrossRefGoogle Scholar
  10. Balazs AB, Chen J, Hong CM et al (2011) Antibody-based protection against HIV infection by vectored immunoprophylaxis. Nature 481:81–84PubMedPubMedCentralCrossRefGoogle Scholar
  11. Bankert RB, Balu-Iyer SV, Odunsi K et al (2011) Humanized mouse model of ovarian cancer recapitulates patient solid tumor progression, ascites formation, and metastasis. PLoS One 6:e24420PubMedPubMedCentralCrossRefGoogle Scholar
  12. Bastide C, Bagnis C, Mannoni P et al (2002) A Nod Scid mouse model to study human prostate cancer. Prostate Cancer Prostatic Dis 5:311–315PubMedCrossRefGoogle Scholar
  13. Bäumler A, Fang FC (2013) Host specificity of bacterial pathogens. Cold Spring Harb Perspect Med 3:a010041PubMedPubMedCentralCrossRefGoogle Scholar
  14. Berges BK, Rowan MR (2011) The utility of the new generation of humanized mice to study HIV-1 infection: Transmission, prevention, pathogenesis, and treatment. Retrovirology 8:65PubMedPubMedCentralCrossRefGoogle Scholar
  15. Beyersdorf N, Kerkau T, Hünig T (2015) CD28 co-stimulation in T-cell homeostasis: A recent perspective. Immunotargets Ther 4:111–122PubMedPubMedCentralGoogle Scholar
  16. Bility MT, Zhang L, Washburn ML et al (2012) Generation of a humanized mouse model with both human immune system and liver cells to model hepatitis C virus infection and liver immunopathogenesis. Nat Protoc 7:1608–1617PubMedPubMedCentralCrossRefGoogle Scholar
  17. Bility MT, Cheng L, Zhang Z et al (2014) Hepatitis B virus infection and immunopathogenesis in a humanized mouse model: Induction of human-specific liver fibrosis and M2-like macrophages. PLoS Pathog 10:e1004032PubMedPubMedCentralCrossRefGoogle Scholar
  18. Billerbeck E, Barry WT, Mu K et al (2011) Development of human CD4(+)FoxP3(+) regulatory T cells in human stem cell factor-, granulocyte-macrophage colony-stimulating factor-, and interleukin-3-expressing NOD-SCID IL2Rγ(null) humanized mice. Blood 117:3076–3086PubMedPubMedCentralCrossRefGoogle Scholar
  19. Bird GA, Polsky A, Estes P et al (2014) Expansion of human and murine hematopoietic stem and progenitor cells ex vivo without genetic modification using MYC and Bcl-2 fusion proteins. PLoS One 9:e105525PubMedPubMedCentralCrossRefGoogle Scholar
  20. Bird BH, Spengler JR, Chakrabarti AK et al (2016) Humanized mouse model of Ebola virus disease mimics the immune responses in human disease. J Infect Dis 213:703–711PubMedCrossRefGoogle Scholar
  21. Blank SV, Rubin SC, Coukos G et al (2002) Replication-selective herpes simplex virus type 1 mutant therapy of cervical cancer is enhanced by low-dose radiation. Hum Gene Ther 13:627–639PubMedCrossRefGoogle Scholar
  22. Blunt T, Gell D, Fox M et al (1996) Identification of a nonsense mutation in the carboxyl-terminal region of DNA-dependent protein kinase catalytic subunit in the scid mouse. Proc Natl Acad Sci USA 93:10285–10290PubMedCrossRefGoogle Scholar
  23. Bosma MJ, Carroll AM (1991) The SCID mouse mutant: Definition, characterization, and potential uses. Annu Rev Immunol 9:323–350PubMedCrossRefGoogle Scholar
  24. Bosma GC, Custer RP, Bosma MJ (1983) A severe combined immunodeficiency mutation in the mouse. Nature 301:527–530PubMedCrossRefGoogle Scholar
  25. Boss IW, Nadeau PE, Abbott JR et al (2011) A Kaposi’s Sarcoma-associated herpesvirus-encoded ortholog of microRNA miR-155 induces human splenic B-cell expansion in NOD/LtSz-scid IL2Rγnull mice. J Virol 85:9877–9886PubMedPubMedCentralCrossRefGoogle Scholar
  26. Brady JL, Harrison LC, Goodman DJ et al (2014) Preclinical screening for acute toxicity of therapeutic monoclonal antibodies in a hu-SCID model. Clin Transl Immunology 3:e29PubMedPubMedCentralCrossRefGoogle Scholar
  27. Brainard DM, Seung E, Frahm N et al (2009) Induction of robust cellular and humoral virus-specific adaptive immune responses in human immunodeficiency virus-infected humanized BLT mice. J Virol 83:7305–7321PubMedPubMedCentralCrossRefGoogle Scholar
  28. Brehm MA, Cuthbert A, Yang C et al (2010) Parameters for establishing humanized mouse models to study human immunity: Analysis of human hematopoietic stem cell engraftment in three immunodeficient strains of mice bearing the IL2rγ(null) mutation. Clin Immunol 135:84–98PubMedPubMedCentralCrossRefGoogle Scholar
  29. Brehm MA, Shultz LD, Luban J et al (2013) Overcoming current limitations in humanized mouse research. J Infect Dis 208(Suppl 2):S125–S130PubMedPubMedCentralCrossRefGoogle Scholar
  30. Calderon VE, Valbuena G, Goez Y et al (2013) A humanized mouse model of Tuberculosis. PLoS One 8:e63331PubMedPubMedCentralCrossRefGoogle Scholar
  31. Carlton JM, Adams JH, Silva JC et al (2008) Comparative genomics of the neglected human malaria parasite Plasmodium vivax. Nature 455:757–763PubMedPubMedCentralCrossRefGoogle Scholar
  32. Carter CC, McNamara LA, Onafuwa-Nuga A et al (2011) HIV-1 utilizes the CXCR4 chemokine receptor to infect multipotent hematopoietic stem and progenitor cells. Cell Host Microbe 9:223–234PubMedPubMedCentralCrossRefGoogle Scholar
  33. Cassidy JW, Caldas C, Bruna A (2015) Maintaining tumour heterogeneity in patient-derived tumour xenografts. Cancer Res 75:2963–2968PubMedPubMedCentralCrossRefGoogle Scholar
  34. Chang H, Wachtman LM, Pearson CB et al (2009) Non-human primate model of Kaposi’s Sarcoma-associated herpesvirus infection. PLoS Pathog 5:e1000606PubMedPubMedCentralCrossRefGoogle Scholar
  35. Chang DK, Moniz RJ, Xu Z et al (2015) Human anti-CAIX antibodies mediate immune cell inhibition of renal cell carcinoma in vitro and in a humanized mouse model in vivo. Mol Cancer 14:119PubMedPubMedCentralCrossRefGoogle Scholar
  36. Chen Q, Khoury M, Chen J (2009) Expression of human cytokines dramatically improves reconstitution of specific human-blood lineage cells in humanized mice. Proc Natl Acad Sci USA 106:21783–21788PubMedCrossRefGoogle Scholar
  37. Chen Q, Kwang J, He F et al (2012) GM-CSF and IL-4 stimulate antibody responses in humanized mice by promoting T, B and dendritic cell maturation. J Immunol 189:5223–5229PubMedPubMedCentralCrossRefGoogle Scholar
  38. Chen Q, Amaladoss A, Ye W et al (2014) Human natural killer cells control Plasmodium falciparum infection by eliminating infected red blood cells. Proc Natl Acad Sci USA 111:1479–1484PubMedCrossRefGoogle Scholar
  39. Chen Q, Ye W, Jian Tan W et al (2015) Delineation of natural killer cell differentiation from myeloid progenitors in human. Sci Rep 5:15118PubMedPubMedCentralCrossRefGoogle Scholar
  40. Choudhary SK, Rezk NL, Ince WL et al (2009) Suppression of human immunodeficiency virus type 1 (HIV-1) viremia with reverse transcriptase and integrase inhibitors, CD4(+) T-cell recovery, and viral rebound upon interruption of therapy in a new model for HIV treatment in the humanized rag2(/)γ(c)(/) mouse. J Virol 83:8254–8258PubMedPubMedCentralCrossRefGoogle Scholar
  41. Choudhary SK, Archin NM, Cheema M et al (2012) Latent HIV-1 infection of resting CD4(+) T cells in the humanized Rag2(/) γ(c)(/) mouse. J Virol 86:114–120PubMedPubMedCentralCrossRefGoogle Scholar
  42. Cocco M, Bellan C, Tussiwand R et al (2008) CD34(+) cord blood cell-transplanted Rag2(/) γ(c)(/) mice as a model for Epstein-Barr virus infection. Am J Pathol 173:1369–1378PubMedPubMedCentralCrossRefGoogle Scholar
  43. Cosgun KN, Rahmig S, Mende N et al (2014) Kit regulates HSC engraftment across the human-mouse species barrier. Cell Stem Cell 15:227–238PubMedCrossRefGoogle Scholar
  44. Covassin L, Laning J, Abdi R et al (2011) Human peripheral blood CD4 T cell-engrafted non-obese diabetic-scid IL2rγ(null) H2-Ab1 (tm1Gru) Tg (human leucocyte antigen D-related 4) mice: A mouse model of human allogeneic graft-versus-host disease. Clin Exp Immunol 166:269–280PubMedPubMedCentralCrossRefGoogle Scholar
  45. Covassin L, Jangalwe S, Jouvet N et al (2013) Human immune system development and survival of non-obese diabetic (NOD)-scid IL2rγnull (NSG) mice engrafted with human thymus and autologous haematopoietic stem cells. Clin Exp Immunol 174:372–388PubMedPubMedCentralCrossRefGoogle Scholar
  46. Daenthanasanmak A, Salguero G, Sundarasetty BS et al (2015) Engineered dendritic cells from cord blood and adult blood accelerate effector T cell immune reconstitution against HCMV. Mol Ther Methods Clin Dev 1:14060PubMedPubMedCentralCrossRefGoogle Scholar
  47. Dash PK, Gorantla S, Gendelman HE et al (2011) Loss of neuronal integrity during progressive HIV-1 infection of humanized mice. J Neurosci 31:3148–3157PubMedPubMedCentralCrossRefGoogle Scholar
  48. Denton PW, Garcia JV (2011) Humanized mouse models of HIV infection. AIDS Rev 13:135–148PubMedPubMedCentralGoogle Scholar
  49. Denton PW, Estes JD, Sun Z et al (2008) Antiretroviral pre-exposure prophylaxis prevents vaginal transmission of HIV-1 in humanized BLT mice. PLoS Med 5:e16PubMedPubMedCentralCrossRefGoogle Scholar
  50. Denton PW, Krisko JF, Powell DA et al (2010) Systemic administration of antiretrovirals prior to exposure prevents rectal and intravenous HIV-1 transmission in humanized BLT mice. PLoS One 5:e8829PubMedPubMedCentralCrossRefGoogle Scholar
  51. Denton PW, Olesen R, Choudhary SK et al (2012) Generation of HIV latency in humanized BLT mice. J Virol 86:630–634PubMedPubMedCentralCrossRefGoogle Scholar
  52. Duchosal MA, Eming SA, McConahey PJ et al (1992) Characterization of hu-PBL-SCID mice with high human immunoglobulin serum levels and graft-versus-host disease. Am J Pathol 141:1097–1113PubMedPubMedCentralGoogle Scholar
  53. Duyne RV, Narayanan A, K-Hall K et al (2011) Humanized mouse models of HIV-1 latency. Curr HIV Res 9:595–605PubMedCrossRefGoogle Scholar
  54. Fauci AS (1988) The human immunodeficiency virus: infectivity and mechanisms of pathogenesis. Science 239:617–622PubMedCrossRefGoogle Scholar
  55. Finnie NJ, Gottlieb TM, Blunt T et al (1996) DNA-dependent protein kinase defects are linked to deficiencies in DNA repair and V(D)J recombination. Philos Trans R Soc Lond B Biol Sci 351:173–179PubMedCrossRefGoogle Scholar
  56. Fischer KE, Austad SN (2011) The development of small primate models for aging research. ILAR J 52:78–88PubMedPubMedCentralCrossRefGoogle Scholar
  57. Fogal B, Yi T, Wang C et al (2011) Neutralizing IL-6 reduces human arterial allograft rejection by allowing emergence of CD161(+) CD4(+) T regulatory cells. J Immunol 187:6268–6280PubMedPubMedCentralCrossRefGoogle Scholar
  58. Frias-Staheli N, Dorner M, Marukian S et al (2014) Utility of humanized BLT mice for analysis of Dengue virus infection and antiviral drug testing. J Virol 88:2205–2218PubMedPubMedCentralCrossRefGoogle Scholar
  59. Fukuchi Y, Miyakawa Y, Kobayashi K et al (1998) Cytokine dependent growth of human TF-1 leukemic cell line in human GM-CSF and IL-3 producing transgenic SCID mice. Leuk Res 22:837–843PubMedCrossRefGoogle Scholar
  60. Geraghty RJ, Capes-Davis A, Davis JM et al (2014) Guidelines for the use of cell lines in biomedical research. Br J Cancer 111:1021–1046PubMedPubMedCentralCrossRefGoogle Scholar
  61. Good MF, Hawkes MT, Yanow SK (2015) Humanized mouse models to study cell-mediated immune responses to liver-stage malaria vaccines. Trends Parasitol 31:583–594PubMedCrossRefGoogle Scholar
  62. Goodman AL, Forbes EK, Williams AR et al (2013) The utility of Plasmodium berghei as a rodent model for anti-merozoite malaria vaccine assessment. Sci Rep 3:1706PubMedPubMedCentralCrossRefGoogle Scholar
  63. Gorantla S, Makarov E, Finke-Dwyer J et al (2010) CD8+ cell depletion accelerates HIV-1 immunopathology in humanized mice. J Immunol 184:7082–7091PubMedPubMedCentralCrossRefGoogle Scholar
  64. Greek R, Rice MJ (2012) Animal models and conserved processes. Theor Biol Med Model 9:40PubMedPubMedCentralCrossRefGoogle Scholar
  65. Grover A, Troy A, Rowe J et al (2017) Humanized NOG mice as a model for tuberculosis vaccine-induced immunity: a comparative analysis with the mouse and guinea pig models of tuberculosis. Immunology 152:150–162PubMedCrossRefPubMedCentralGoogle Scholar
  66. Gunawan M, Her ZS, Liu M et al (2017) A novel human systemic lupus erythematosus model in humanised mice. Sci Rep 7:16642PubMedPubMedCentralCrossRefGoogle Scholar
  67. Harris DT, Badowski M (2014) Long term human reconstitution and immune aging in NOD-Rag ()-γ chain () mice. Immunobiology 219:131–137PubMedCrossRefGoogle Scholar
  68. Harris DT, Badowski M, Balamurugan A et al (2013) Long-term human immune system reconstitution in non-obese diabetic (NOD)-Rag ()-γ chain () (NRG) mice is similar but not identical to the original stem cell donor. Clin Exp Immunol 174:402–413PubMedPubMedCentralCrossRefGoogle Scholar
  69. Harui A, Kiertscher SM, Roth MD (2011) Reconstitution of huPBL-NSG mice with donor-matched dendritic cells enables antigen-specific T-cell activation. J Neuroimmune Pharmacol 6:148–157PubMedCrossRefGoogle Scholar
  70. Hatziioannou T, Evans DT (2012) Animal models for HIV/AIDS research. Nat Rev Microbiol 10:852–867PubMedPubMedCentralCrossRefGoogle Scholar
  71. Her Z, Yong KS, Paramasivam K et al (2017) An improved pre-clinical patient-derived liquid xenograft mouse model for acute myeloid leukemia. J Hematol Oncol 10:162PubMedPubMedCentralCrossRefGoogle Scholar
  72. Holt N, Wang J, Kim K et al (2010) Zinc finger nuclease-mediated CCR5 knockout hematopoietic stem cell transplantation controls HIV-1 in vivo. Nat Biotechnol 28:839–847PubMedPubMedCentralCrossRefGoogle Scholar
  73. Horvath C, Andrews L, Baumann A et al (2012) Storm forecasting: additional lessons from the CD28 superagonist TGN1412 trial. Nat Rev Immunol 12:740PubMedGoogle Scholar
  74. Hu Z, Yang YG (2012) Full reconstitution of human platelets in humanized mice after macrophage depletion. Blood 120:1713–1716PubMedPubMedCentralCrossRefGoogle Scholar
  75. Hu Z, Van Rooijen N, Yang YG (2011) Macrophages prevent human red blood cell reconstitution in immunodeficient mice. Blood 118:5938–5946PubMedPubMedCentralCrossRefGoogle Scholar
  76. Huntington ND, Legrand N, Alves NL et al (2009) IL-15 trans-presentation promotes human NK cell development and differentiation in vivo. J Exp Med 206:25–34PubMedPubMedCentralCrossRefGoogle Scholar
  77. Ince WL, Zhang L, Jiang Q et al (2010) Evolution of the HIV-1 env gene in the Rag2(/) γ(C)(/) humanized mouse model. J Virol 84:2740–2752PubMedCrossRefGoogle Scholar
  78. Ishikawa F, Yasukawa M, Lyons B et al (2005) Development of functional human blood and immune systems in NOD/SCID/IL2 receptor γ chain(null) mice. Blood 106:1565–1573PubMedPubMedCentralCrossRefGoogle Scholar
  79. Ito M, Hiramatsu H, Kobayashi K et al (2002) NOD/SCID/gamma(c)(null) mouse: an excellent recipient mouse model for engraftment of human cells. Blood 100:3175–3182PubMedCrossRefGoogle Scholar
  80. Ito A, Ishida T, Yano H et al (2009) Defucosylated anti-CCR4 monoclonal antibody exercises potent ADCC-mediated antitumor effect in the novel tumor-bearing humanized NOD/Shi-scid, IL-2Rγnull mouse model. Cancer Immunol Immunother 58:1195–1206PubMedCrossRefGoogle Scholar
  81. Ito R, Takahashi T, Katano I et al (2013) Establishment of a human allergy model using human IL-3/GM-CSF—transgenic NOG mice. J Immunol 191:2890–2899PubMedCrossRefGoogle Scholar
  82. Jangalwe S, Shultz LD, Mathew A et al (2016) Improved B cell development in humanized NOD-scid IL2Rγnull mice transgenically expressing human stem cell factor, granulocyte-macrophage colony-stimulating factor and interleukin-3. Immun Inflamm Dis 4:427–440PubMedPubMedCentralCrossRefGoogle Scholar
  83. Jiménez-Díaz MB, Mulet T, Viera S et al (2009) Improved murine model of malaria using Plasmodium falciparum competent strains and non-myelodepleted NOD-scid IL2Rγ(null) mice engrafted with human erythrocytes. Antimicrob Agents Chemother 53:4533–4536PubMedPubMedCentralCrossRefGoogle Scholar
  84. Joseph A, Zheng JH, Chen K et al (2010) Inhibition of in vivo HIV infection in humanized mice by gene therapy of human hematopoietic stem cells with a lentiviral vector encoding a broadly neutralizing anti-HIV antibody. J Virol 84:6645–6653PubMedPubMedCentralCrossRefGoogle Scholar
  85. Kapp MB (2006) Ethical and legal issues in research involving human subjects: Do you want a piece of me? J Clin Pathol 59:335–339PubMedPubMedCentralCrossRefGoogle Scholar
  86. Katano I, Ito R, Kamisako T et al (2014) NOD-Rag2(null) IL-2Rγ(null) mice: An alternative to NOG mice for generation of humanized mice. Exp Anim 63:321–330PubMedPubMedCentralCrossRefGoogle Scholar
  87. Keng CT, Sze CW, Zheng D et al (2015) Characterisation of liver pathogenesis, human immune responses and drug testing in a humanised mouse model of HCV infection. Gut 65:1744–1753PubMedPubMedCentralCrossRefGoogle Scholar
  88. King M, Pearson T, Shultz LD et al (2008) A new Hu-PBL model for the study of human islet alloreactivity based on NOD-scid mice bearing a targeted mutation in the IL-2 receptor gamma chain gene. Clin Immunol 126:303–314PubMedCrossRefGoogle Scholar
  89. Kirkiles-Smith NC, Harding MJ, Shepherd BR et al (2009) Development of a humanized mouse model to study the role of macrophages in allograft injury. Transplantation 87:189–197PubMedPubMedCentralCrossRefGoogle Scholar
  90. Koboziev I, Jones-Hall Y, Valentine JF et al (2015) Use of humanized mice to study the pathogenesis of autoimmune and inflammatory diseases. Inflamm Bowel Dis 21:1652–1673PubMedPubMedCentralCrossRefGoogle Scholar
  91. Kumar P, Ban HS, Kim SS et al (2008) T cell-specific siRNA delivery suppresses HIV-1 infection in humanized mice. Cell 134:577–586PubMedPubMedCentralCrossRefGoogle Scholar
  92. Kuruvilla JG, Troyer RM, Devi S et al (2007) Dengue virus infection and immune response in humanized RAG2/γc/ (RAG-hu) mice. Virology 369:143–152PubMedCrossRefGoogle Scholar
  93. Lan P, Wang L, Diouf B et al (2004) Induction of human T-cell tolerance to porcine xenoantigens through mixed hematopoietic chimerism. Blood 103:3964–3969PubMedCrossRefGoogle Scholar
  94. Lan P, Tonomura N, Shimizu A et al (2006) Reconstitution of a functional human immune system in immunodeficient mice through combined human fetal thymus/liver and CD34+ cell transplantation. Blood 108:487–492PubMedCrossRefGoogle Scholar
  95. Lang J, Kelly M, Freed BM et al (2013) Studies of lymphocyte reconstitution in a humanized mouse model reveal a requirement of T cells for human B cell maturation. J Immunol 190:2090–2101PubMedPubMedCentralCrossRefGoogle Scholar
  96. Li G, Cheng M, Nunoya J et al (2014) Plasmacytoid dendritic cells suppress HIV-1 replication but contribute to HIV-1 induced immunopathogenesis in humanized mice. PLoS Pathog 10:e1004291PubMedPubMedCentralCrossRefGoogle Scholar
  97. Lieber MR, Hesse JE, Lewis S et al (1988) The defect in murine severe combined immune deficiency: joining of signal sequences but not coding segments in V(D)J recombination. Cell 55:7–16PubMedCrossRefGoogle Scholar
  98. Lim AI, Li Y, Lopez-Lastra S et al (2017) Systemic human ILC precursors provide a substrate for tissue ILC differentiation. Cell 168:1086–1100.e10PubMedCrossRefGoogle Scholar
  99. Long BR, Stoddart CA (2012) Alpha interferon and HIV infection cause activation of human T cells in NSG-BLT mice. J Virol 86:3327–3336PubMedPubMedCentralCrossRefGoogle Scholar
  100. Lüdtke A, Oestereich L, Ruibal P et al (2015) Ebola virus disease in mice with transplanted human hematopoietic stem cells. J Virol 89:4700–4704PubMedPubMedCentralCrossRefGoogle Scholar
  101. Marsden MD, Kovochich M, Suree N et al (2012) HIV latency in the humanized BLT mouse. J Virol 86:339–347PubMedPubMedCentralCrossRefGoogle Scholar
  102. Maykel J, Liu JH, Li H et al (2014) NOD-scidIl2rg(tm1Wjl) and NOD-Rag1(null)Il2rg(tm1Wjl): A model for stromal cell–tumor cell interaction for human colon cancer. Dig Dis Sci 59:1169–1179PubMedPubMedCentralCrossRefGoogle Scholar
  103. McKenzie R, Fried MW, Sallie R et al (1995) Hepatic failure and lactic acidosis due to Fialuridine (FIAU), an investigational nucleoside analogue for chronic hepatitis B. N Engl J Med 333:1099–1105PubMedCrossRefGoogle Scholar
  104. Melkus MW, Estes JD, Padgett-Thomas A et al (2006) Humanized mice mount specific adaptive and innate immune responses to EBV and TSST-1. Nat Med 12:1316–1322PubMedCrossRefGoogle Scholar
  105. Mestas J, Hughes CC (2004) Of mice and not men: Differences between mouse and human immunology. J Immunol 172:2731–2738PubMedCrossRefGoogle Scholar
  106. Miller RJ, Cairns JS, Bridges S et al (2000) Human immunodeficiency virus and AIDS: Insights from animal lentiviruses. J Virol 74:7187–7195PubMedPubMedCentralCrossRefGoogle Scholar
  107. Miyakawa Y, Ohnishi Y, Tomisawa M et al (2004) Establishment of a new model of human multiple myeloma using NOD/SCID/γcnull (NOG) mice. Biochem Biophys Res Commun 313:258–262PubMedCrossRefGoogle Scholar
  108. Moffat JF, Stein MD, Kaneshima H et al (1995) Tropism of varicella-zoster virus for human CD4+ and CD8+ T lymphocytes and epidermal cells in SCID-hu mice. J Virol 69:5236–5242PubMedPubMedCentralGoogle Scholar
  109. Morton JJ, Bird G, Keysar SB et al (2016) XactMice: humanizing mouse bone marrow enables microenvironment reconstitution in a patient-derived xenograft model of head and neck cancer. Oncogene 35:290–300PubMedCrossRefGoogle Scholar
  110. Mota J, Rico-Hesse R (2009) Humanized mice show clinical signs of dengue fever according to infecting virus genotype. J Virol 83:8638–8645PubMedPubMedCentralCrossRefGoogle Scholar
  111. Mota J, Rico-Hesse R (2011) Dengue virus tropism in humanized mice recapitulates human dengue fever. PLoS One 6:e20762PubMedPubMedCentralCrossRefGoogle Scholar
  112. Neff CP, Zhou J, Remling L et al (2011) An aptamer-siRNA chimera suppresses HIV-1 viral loads and protects from helper CD4(+) T cell decline in humanized mice. Sci Transl Med 3:66ra6PubMedPubMedCentralCrossRefGoogle Scholar
  113. Nicolini FE, Cashman JD, Hogge DE et al (2004) NOD//SCID mice engineered to express human IL-3, GM-CSF and Steel factor constitutively mobilize engrafted human progenitors and compromise human stem cell regeneration. Leukemia 18:341–347PubMedCrossRefGoogle Scholar
  114. Nusbaum RJ, Calderon VE, Huante MB et al (2016) Pulmonary tuberculosis in humanized mice infected with HIV-1. Sci Rep 6:21522PubMedPubMedCentralCrossRefGoogle Scholar
  115. Onoe T, Kalscheuer H, Danzl N et al (2011) Human natural regulatory T cell development, suppressive function and post-thymic maturation in a humanized mouse model. J Immunol 187:3895–3903PubMedPubMedCentralCrossRefGoogle Scholar
  116. Oosting M, Cheng SC, Bolscher JM et al (2014) Human TLR10 is an anti-inflammatory pattern-recognition receptor. Proc Natl Acad Sci USA 111:E4478–E4484PubMedCrossRefGoogle Scholar
  117. Pain A, Böhme U, Berry AE et al (2008) The genome of the simian and human malaria parasite Plasmodium knowlesi. Nature 455:799–803PubMedPubMedCentralCrossRefGoogle Scholar
  118. Pan CX, Shi W, Ma AH et al (2017) Humanized mice (humice) carrying patient-derived xenograft (PDX) as a platform to develop immunotherapy in bladder cancer (BCa). J Clin Oncol 35:381CrossRefGoogle Scholar
  119. Patton J, Vuyyuru R, Siglin A et al (2015) Evaluation of the efficiency of human immune system reconstitution in NSG mice and NSG mice containing a human HLA.A2 transgene using hematopoietic stem cells purified from different sources. J Immunol Methods 422:13–21PubMedPubMedCentralCrossRefGoogle Scholar
  120. Pearson T, Greiner DL, Shultz LD (2008) Creation of “humanized” mice to study human immunity. Curr Protoc Immunol Chap 15:Unit-15.21Google Scholar
  121. Phillips KA, Bales KL, Capitanio JP et al (2014) Why primate models matter. Am J Primatol 76:801–827PubMedPubMedCentralCrossRefGoogle Scholar
  122. Ploss A, Evans MJ, Gaysinskaya VA et al (2009) Human occludin is a hepatitis C virus entry factor required for infection of mouse cells. Nature 457:882–886PubMedPubMedCentralCrossRefGoogle Scholar
  123. Rahmig S, Kronstein-Wiedemann R, Fohgrub J et al (2016) Improved human erythropoiesis and platelet formation in humanized NSGW41 mice. Stem Cell reports 7:591–601PubMedPubMedCentralCrossRefGoogle Scholar
  124. Rehman W, Arfons LM, Lazarus HM (2011) The rise, fall and subsequent triumph of Thalidomide: Lessons learned in drug development. Ther Adv Hematol 2:291–308PubMedPubMedCentralCrossRefGoogle Scholar
  125. Rodríguez E, Ip WH, Kolbe V et al (2017) Humanized mice reproduce acute and persistent human adenovirus infection. J Infect Dis 215:70–79PubMedCrossRefGoogle Scholar
  126. Rongvaux A, Willinger T, Takizawa H et al (2011) Human thrombopoietin knockin mice efficiently support human hematopoiesis in vivo. Proc Natl Acad Sci USA 108:2378–2383PubMedCrossRefGoogle Scholar
  127. Rongvaux A, Willinger T, Martinek J et al (2014) Development and function of human innate immune cells in a humanized mouse model. Nat Biotechnol 32:364–372PubMedPubMedCentralCrossRefGoogle Scholar
  128. Roth MD, Harui A (2015) Human tumor infiltrating lymphocytes cooperatively regulate prostate tumor growth in a humanized mouse model. J Immunother Cancer 3:12PubMedPubMedCentralCrossRefGoogle Scholar
  129. Sanmamed MF, Rodriguez I, Schalper KA et al (2015) Nivolumab and Urelumab enhance antitumor activity of human T lymphocytes engrafted in Rag2/IL2Rγnull immunodeficient mice. Cancer Res 75:3466–3478PubMedCrossRefGoogle Scholar
  130. Sato K, Nie C, Misawa N et al (2010) Dynamics of memory and naïve CD8+ T lymphocytes in humanized NOD/SCID/IL-2Rγnull mice infected with CCR5-tropic HIV-1. Vaccine 28(Suppl 2):B32–B37PubMedCrossRefGoogle Scholar
  131. Sato K, Misawa N, Nie C et al (2011) A novel animal model of Epstein-Barr virus—associated hemophagocytic lymphohistiocytosis in humanized mice. Blood 117:5663–5673PubMedCrossRefGoogle Scholar
  132. Schneider M, Ekholm F, Grönvik KO (1997) Severe graft-versus-host disease in SCID mice is associated with a decrease of selective donor cell TCR Vβ specificities and increased expression of IFN-γ and IL-4. Scand J Immunol 46:147–158PubMedCrossRefGoogle Scholar
  133. Shanks N, Greek R, Greek J (2009) Are animal models predictive for humans? Philos Ethics Humanit Med 4:2PubMedPubMedCentralCrossRefGoogle Scholar
  134. Sheng-Tanner X, McKerlie C, Spaner D (2000) Characterization of graft-versus-host disease in scid mice and prevention by physicochemical stressors. Transplantation 70:1683–1693PubMedCrossRefGoogle Scholar
  135. Shi Y, Inoue H, Wu JC et al (2017) Induced pluripotent stem cell technology: a decade of progress. Nat Rev Drug Discov 16:115–130PubMedCrossRefGoogle Scholar
  136. Shimizu S, Hong P, Arumugam B et al (2010) A highly efficient short hairpin RNA potently down-regulates CCR5 expression in systemic lymphoid organs in the hu-BLT mouse model. Blood 115:1534–1544PubMedPubMedCentralCrossRefGoogle Scholar
  137. Shultz LD, Lyons BL, Burzenski LM et al (2005) Human lymphoid and myeloid cell development in NOD/LtSz-scid IL2Rγnull mice engrafted with mobilized human hemopoietic stem cells. J Immunol 174:6477–6489PubMedCrossRefGoogle Scholar
  138. Shultz LD, Pearson T, King M et al (2007) Humanized NOD/LtSz-scid IL2 receptor common gamma chain knockout mice in diabetes research. Ann NY Acad Sci 1103:77–89PubMedCrossRefGoogle Scholar
  139. Shultz LD, Saito Y, Najima Y et al (2010) Generation of functional human T-cell subsets with HLA-restricted immune responses in HLA class I expressing NOD/SCID/IL2rγnull humanized mice. Proc Natl Acad Sci USA 107:13022–13027PubMedCrossRefGoogle Scholar
  140. Shultz LD, Brehm MA, Garcia JV et al (2012) Humanized mice for immune system investigation: Progress, promise and challenges. Nat Rev Immunol 12:786–798PubMedPubMedCentralCrossRefGoogle Scholar
  141. Simpson-Abelson MR, Sonnenberg GF, Takita H et al (2008) Long-term engraftment and expansion of tumor-derived memory T cells following the implantation of non-disrupted pieces of human lung tumor into NOD-scid IL2Rγnull mice. J Immunol 180:7009–7018PubMedCrossRefGoogle Scholar
  142. Smith MS, Goldman DC, Bailey AS et al (2010) G-CSF reactivates human cytomegalovirus in a latently infected humanized mouse model. Cell Host Microbe 8:284–291PubMedPubMedCentralCrossRefGoogle Scholar
  143. Soulard V, Bosson-Vanga H, Lorthiois A et al (2015) Plasmodium falciparum full life cycle and Plasmodium ovale liver stages in humanized mice. Nat Commun 6:7690PubMedPubMedCentralCrossRefGoogle Scholar
  144. Sridharan A, Chen Q, Tang KF et al (2013) Inhibition of megakaryocyte development in the bone marrow underlies Dengue virus-induced thrombocytopenia in humanized mice. J Virol 87:11648–11658PubMedPubMedCentralCrossRefGoogle Scholar
  145. Stoddart CA, Galkina SA, Joshi P et al (2014) Efficacy of broadly neutralizing monoclonal antibody PG16 in HIV-infected humanized mice. Virology 462–463:115–125PubMedCrossRefGoogle Scholar
  146. Strick-Marchand H, Dusséaux M, Darche S et al (2015) A novel mouse model for stable engraftment of a human immune system and human hepatocytes. PLoS One 10:e0119820PubMedPubMedCentralCrossRefGoogle Scholar
  147. Subramanya S, Kim S-S, Abraham S et al (2010) Targeted delivery of small interfering RNA to human dendritic cells to suppress dengue virus infection and associated proinflammatory cytokine production. J Virol 84:2490–2501PubMedCrossRefGoogle Scholar
  148. Sun Z, Denton PW, Estes JD et al (2007) Intrarectal transmission, systemic infection, and CD4(+) T cell depletion in humanized mice infected with HIV-1. J Exp Med 204:705–714PubMedPubMedCentralCrossRefGoogle Scholar
  149. Sun H, Tsai Y, Nowak I et al (2012) Eltrombopag, a thrombopoietin receptor agonist, enhances human umbilical cord blood hematopoietic stem/primitive progenitor cell expansion and promotes multi-lineage hematopoiesis. Stem Cell Res 9:77–86PubMedPubMedCentralCrossRefGoogle Scholar
  150. Taccioli GE, Amatucci AG, Beamish HJ et al (1998) Targeted disruption of the catalytic subunit of the DNA-PK gene in mice confers severe combined immunodeficiency and radiosensitivity. Immunity 9:355–366PubMedCrossRefGoogle Scholar
  151. Tan-Garcia A, Wai L-E, Zheng D et al (2017) Intrahepatic CD206+ macrophages contribute to inflammation in advanced viral-related liver disease. J Hepatol 67:490–500PubMedCrossRefGoogle Scholar
  152. Tary-Lehmann M, Saxon A, Lehmann PV (1995) The human immune system in hu-PBL-SCID mice. Immunol Today 16:529–533PubMedCrossRefGoogle Scholar
  153. Tezuka K, Xun R, Tei M et al (2014) An animal model of adult T-cell leukemia: Humanized mice with HTLV-1—specific immunity. Blood 123:346–355PubMedCrossRefGoogle Scholar
  154. Tobin LM, Healy ME, English K et al (2013) Human mesenchymal stem cells suppress donor CD4+ T cell proliferation and reduce pathology in a humanized mouse model of acute graft-versus-host disease. Clin Exp Immunol 172:333–348PubMedPubMedCentralCrossRefGoogle Scholar
  155. Tonomura N, Habiro K, Shimizu A et al (2008) Antigen-specific human T-cell responses and T cell–dependent production of human antibodies in a humanized mouse model. Blood 111:4293–4296PubMedPubMedCentralCrossRefGoogle Scholar
  156. Traggiai E, Chicha L, Mazzucchelli L et al (2004) Development of a human adaptive immune system in cord blood cell-transplanted mice. Science 304:104–107PubMedCrossRefGoogle Scholar
  157. Tsoneva D, Minev B, Frentzen A et al (2017) Humanized mice with subcutaneous human solid tumors for immune response analysis of vaccinia virus-mediated oncolysis. Mol Ther Oncolytics 5:41–61PubMedPubMedCentralCrossRefGoogle Scholar
  158. Tsuji M, Ishihara C, Arai S et al (1995) Establishment of a SCID mouse model having circulating human red blood cells and a possible growth of Plasmodium falciparum in the mouse. Vaccine 13:1389–1392PubMedCrossRefGoogle Scholar
  159. Unger WW, Pearson T, Abreu JR et al (2012) Islet-specific CTL cloned from a type 1 diabetes patient cause beta-cell destruction after engraftment into HLA-A2 transgenic NOD/SCID/IL2RG null mice. PLoS One 7:e49213PubMedPubMedCentralCrossRefGoogle Scholar
  160. Valbuena G, Halliday H, Borisevich V et al (2014) A human lung xenograft mouse model of Nipah virus infection. PLoS Pathog 10:e1004063PubMedPubMedCentralCrossRefGoogle Scholar
  161. Van Lent AU, Dontje W, Nagasawa M et al (2009) IL-7 enhances thymic human T cell development in “human immune system” Rag2/IL-2Rγc/ mice without affecting peripheral T cell homeostasis. J Immunol 183:7645–7655PubMedCrossRefGoogle Scholar
  162. Van der Loo JC, Hanenberg H, Cooper RJ et al (1998) Nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mouse as a model system to study the engraftment and mobilization of human peripheral blood stem cells. Blood 92:2556–2570PubMedGoogle Scholar
  163. Van der Worp HB, Howells DW, Sena ES et al (2010) Can animal models of disease reliably inform human studies? PLoS Med 7:e1000245PubMedPubMedCentralCrossRefGoogle Scholar
  164. Vanden Haesevelde MM, Peeters M, Jannes G et al (1996) Sequence analysis of a highly divergent HIV-1-related lentivirus isolated from a wild captured chimpanzee. Virology 221:346–350PubMedCrossRefGoogle Scholar
  165. Vaughan AM, Mikolajczak SA, Wilson EM et al (2012) Complete Plasmodium falciparum liver-stage development in liver-chimeric mice. J Clin Invest 122:3618–3628PubMedPubMedCentralCrossRefGoogle Scholar
  166. Viehmann Milam AA, Maher SE, Gibson JA et al (2014) A humanized mouse model of autoimmune insulitis. Diabetes 63:1712–1724PubMedPubMedCentralCrossRefGoogle Scholar
  167. Vuyyuru R, Liu H, Manser T et al (2011) Characteristics of Borrelia hermsii infection in human hematopoietic stem cell-engrafted mice mirror those of human relapsing fever. Proc Natl Acad Sci USA 108:20707–20712PubMedCrossRefGoogle Scholar
  168. Waldron-Lynch F, Deng S, Preston-Hurlburt P et al (2012) Analysis of human biologics with a mouse skin transplant model in humanized mice. Am J Transplant 12:2652–2662PubMedCrossRefGoogle Scholar
  169. Wang X, Berger C, Wong CW et al (2011) Engraftment of human central memory-derived effector CD8(+) T cells in immunodeficient mice. Blood 117:1888–1898PubMedPubMedCentralCrossRefGoogle Scholar
  170. Wang LX, Kang G, Kumar P et al (2014) Humanized-BLT mouse model of Kaposi’s sarcoma-associated herpesvirus infection. Proc Natl Acad Sci USA 111:3146–3151PubMedCrossRefGoogle Scholar
  171. Washburn ML, Bility MT, Zhang L et al (2011) A humanized mouse model to study hepatitis C virus infection, immune response, and liver disease. Gastroenterol 140:1334–1344CrossRefGoogle Scholar
  172. Watanabe S, Ohta S, Yajima M et al (2007) Humanized NOD/SCID/IL2Rγnull mice transplanted with hematopoietic stem cells under nonmyeloablative conditions show prolonged life spans and allow detailed analysis of human immunodeficiency virus type 1 pathogenesis. J Virol 81:13259–13264PubMedPubMedCentralCrossRefGoogle Scholar
  173. Watanabe Y, Takahashi T, Okajima A et al (2009) The analysis of the functions of human B and T cells in humanized NOD/shi-scid/γcnull (NOG) mice (hu-HSC NOG mice). Int Immunol 21:843–858PubMedCrossRefGoogle Scholar
  174. Wege AK, Ernst W, Eckl J et al (2011) Humanized tumor mice—a new model to study and manipulate the immune response in advanced cancer therapy. Int J Cancer 129:2194–2206PubMedCrossRefGoogle Scholar
  175. Wege AK, Florian C, Ernst W et al (2012) Leishmania major infection in humanized mice induces systemic infection and provokes a nonprotective human immune response. PLoS Negl Trop Dis 6:e1741PubMedPubMedCentralCrossRefGoogle Scholar
  176. Wege AK, Schmidt M, Ueberham E et al (2014) Co-transplantation of human hematopoietic stem cells and human breast cancer cells in NSG mice: A novel approach to generate tumor cell specific human antibodies. MAbs 6:968–977PubMedPubMedCentralCrossRefGoogle Scholar
  177. Weißmüller S, Kronhart S, Kreuz D et al (2016) TGN1412 induces lymphopenia and human cytokine release in a humanized mouse model. PLoS One 11:e0149093PubMedPubMedCentralCrossRefGoogle Scholar
  178. Whitfield-Larry F, Young EF, Talmage G et al (2011) HLA-A2–matched peripheral blood mononuclear cells from type 1 diabetic patients, but not nondiabetic donors, transfer insulitis to NOD-scid/γc(null)/HLA-A2 transgenic mice concurrent with the expansion of islet-specific CD8(+) T cells. Diabetes 60:1726–1733PubMedPubMedCentralCrossRefGoogle Scholar
  179. Xia Z, Taylor PR, Locklin RM et al (2006) Innate immune response to human bone marrow fibroblastic cell implantation in CB17 scid/beige mice. J Cell Biochem 98:966–980PubMedCrossRefGoogle Scholar
  180. Xu D, Nishimura T, Nishimura S et al (2014) Fialuridine induces acute liver failure in chimeric TK-NOG mice: A model for detecting hepatic drug toxicity prior to human testing. PLoS Med 11:e1001628PubMedPubMedCentralCrossRefGoogle Scholar
  181. Yajima M, Imadome K, Nakagawa A et al (2008) A new humanized mouse model of Epstein-Barr virus infection that reproduces persistent infection, lymphoproliferative disorder, and cell-mediated and humoral immune responses. J Infect Dis 198:673–682PubMedCrossRefGoogle Scholar
  182. Yamada E, Yoshikawa R, Nakano Y et al (2015) Impacts of humanized mouse models on the investigation of HIV-1 infection: Illuminating the roles of viral accessory proteins in vivo. Viruses 7:1373–1390PubMedPubMedCentralCrossRefGoogle Scholar
  183. Yao LC, Cheng M, Wang M et al (2016) Patient-derived tumor xenografts in humanized NSG-SGM3 mice: A new immuno-oncology platform. Eur J Cancer 61:S203–S204CrossRefGoogle Scholar
  184. Yong KSM, Keng CT, Tan SQ et al (2016) Human CD34loCD133lo fetal liver cells support the expansion of human CD34hiCD133hi hematopoietic stem cells. Cell Mol Immunol 13:605–614PubMedCrossRefGoogle Scholar
  185. Young NA, Wu LC, Bruss M et al (2015) A chimeric human–mouse model of Sjögren’s syndrome. Clin Immunol 156:1–8PubMedCrossRefGoogle Scholar
  186. Yu CI, Gallegos M, Marches F et al (2008) Broad influenza-specific CD8(+) T-cell responses in humanized mice vaccinated with influenza virus vaccines. Blood 112:3671–3678PubMedPubMedCentralCrossRefGoogle Scholar
  187. Zayoud M, El Malki K, Frauenknecht K et al (2013) Subclinical CNS inflammation as response to a myelin antigen in humanized mice. J Neuroimmune Pharmacol 8:1037–1047PubMedCrossRefGoogle Scholar
  188. Zhang L, Jiang Q, Li G et al (2011) Efficient infection, activation, and impairment of pDCs in the BM and peripheral lymphoid organs during early HIV-1 infection in humanized rag2(/)γ C(/) mice in vivo. Blood 117:6184–6192PubMedPubMedCentralCrossRefGoogle Scholar
  189. Zhang T, Zhang L, Fan S et al (2015) Patient-derived gastric carcinoma xenograft mouse models faithfully represent human tumor molecular diversity. PLoS One 10:e0134493PubMedPubMedCentralCrossRefGoogle Scholar
  190. Zhao T, Zhang Z-n, Westenskow PD et al (2015) Humanized mice reveal differential immunogenicity of cells derived from autologous induced pluripotent stem cells. Cell Stem Cell 17:353–359PubMedCrossRefGoogle Scholar
  191. Zheng J, Wu WL, Liu Y et al (2015) The therapeutic effect of pamidronate on lethal avian influenza A H7N9 virus infected humanized mice. PLoS One 10:e0135999PubMedPubMedCentralCrossRefGoogle Scholar

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© The Author(s) 2018

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Authors and Affiliations

  • Kylie Su Mei Yong
    • 1
    • 2
  • Zhisheng Her
    • 1
  • Qingfeng Chen
    • 1
    • 3
    • 4
    Email author
  1. 1.Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A*STAR)SingaporeSingapore
  2. 2.Department of Biochemistry, Yong Loo Lin School of MedicineNational University of SingaporeSingaporeSingapore
  3. 3.Department of Microbiology and Immunology, Yong Loo Lin School of MedicineNational University of SingaporeSingaporeSingapore
  4. 4.Key Laboratory for Major Obstetric Diseases of Guangdong Province, The Third Affiliated Hospital of Guangzhou Medical UniversityGuangzhouChina

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