Development of an Inflammatory CD14+ Dendritic Cell Subset in Humanized Mice

Humanized mouse models are attractive experimental models for analyzing the development and functions of human dendritic cells (DCs) in vivo. Although various types of DC subsets, including DC type 3 (DC3s), have been identified in humans, it remains unclear whether humanized mice can reproduce heterogeneous DC subsets. CD14, classically known as a monocyte/macrophage marker, is reported as an indicator of DC3s. We previously observed that some CD14+ myeloid cells expressed CD1c, a pan marker for bona fide conventional DC2 (cDC2s), in humanized mouse models in which human FLT3L and GM-CSF genes were transiently expressed using in vivo transfection (IVT). Here, we aimed to elucidate the identity of CD14+CD1c+ DC-like cells in humanized mouse models. We found that CD14+CD1c+ cells were phenotypically different from cDC2s; CD14+CD1c+ cells expressed CD163 but not CD5, whereas cDC2s expressed CD5 but not CD163. Furthermore, CD14+CD1c+ cells primed and polarized naïve CD4+ T cells toward IFN-γ+ Th1 cells more profoundly than cDC2s. Transcriptional analysis revealed that CD14+CD1c+ cells expressed several DC3-specific transcripts, such as CD163, S100A8, and S100A9, and were clearly segregated from cDC2s and monocytes. When lipopolysaccharide was administered to the humanized mice, the frequency of CD14+CD1c+ cells producing IL-6 and TNF-α was elevated, indicating a pro-inflammatory signature. Thus, humanized mice are able to sustain development of functional CD14+CD1c+ DCs, which are equivalent to DC3 subset observed in humans, and they could be useful for analyzing the development and function of DC3s in vivo.

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Perspectives on Non-BLT Humanized Mouse Models for Studying HIV Pathogenesis and Therapy

Viruses ◽

10.3390/v13050776 ◽

2021 ◽

Vol 13 (5) ◽

pp. 776

Author(s):

Kazutaka Terahara ◽

Ryutaro Iwabuchi ◽

Yasuko Tsunetsugu-Yokota

Keyword(s):

Mouse Models ◽

Humanized Mice ◽

Lymphoid Tissues ◽

Humanized Mouse ◽

Hiv Pathogenesis ◽

Hematopoietic Stem ◽

Critical Issues ◽

In Vivo Animal Models ◽

Humanized Mouse Models

A variety of humanized mice, which are reconstituted only with human hematopoietic stem cells (HSC) or with fetal thymus and HSCs, have been developed and widely utilized as in vivo animal models of HIV-1 infection. The models represent some aspects of HIV-mediated pathogenesis in humans and are useful for the evaluation of therapeutic regimens. However, there are several limitations in these models, including their incomplete immune responses and poor distribution of human cells to the secondary lymphoid tissues. These limitations are common in many humanized mouse models and are critical issues that need to be addressed. As distinct defects exist in each model, we need to be cautious about the experimental design and interpretation of the outcomes obtained using humanized mice. Considering this point, we mainly characterize the current conventional humanized mouse reconstituted only with HSCs and describe past achievements in this area, as well as the potential contributions of the humanized mouse models for the study of HIV pathogenesis and therapy. We also discuss the use of various technologies to solve the current problems. Humanized mice will contribute not only to the pre-clinical evaluation of anti-HIV regimens, but also to a deeper understanding of basic aspects of HIV biology.

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Generation of Novel Human Red Blood Cell-Bearing Humanized Mouse Models Based on C3-Deficient NOG Mice

Frontiers in Immunology ◽

10.3389/fimmu.2021.671648 ◽

2021 ◽

Vol 12 ◽

Author(s):

Takuya Yamaguchi ◽

Ikumi Katano ◽

Iyo Otsuka ◽

Ryoji Ito ◽

Misa Mochizuki ◽

...

Keyword(s):

Mouse Models ◽

Complement C3 ◽

Humanized Mouse ◽

Hematopoietic Stem ◽

Gm Csf ◽

Human Red Blood Cells ◽

Mouse Macrophages ◽

C3 Gene ◽

Humanized Mouse Models

Despite recent advances in immunodeficient mouse models bearing human red blood cells (hRBCs), the elimination of circulating hRBCs by residual innate immune systems remains a significant challenge. In this study, we evaluated the role of mouse complement C3 in the elimination of circulating hRBCs by developing a novel NOG substrain harboring a truncated version of the murine C3 gene (NOG-C3ΔMG2-3). Genetic C3 deletion prolonged the survival of transfused hRBCs in the circulation. Chemical depletion and functional impairment of mouse macrophages, using clodronate liposomes (Clo-lip) or gadolinium chloride (GdCl3), respectively, further extended the survival of hRBCs in NOG-C3ΔMG2-3 mice. Low GdCl3 toxicity allowed the establishment of hRBC-bearing mice, in which hRBCs survived for more than 4 weeks with transfusion once a week. In addition, erythropoiesis of human hematopoietic stem cells (hHSCs) was possible in NOG-C3ΔMG2-3/human GM-CSF-IL-3 transgenic mice with Clo-lip treatment. These findings indicate that mouse models harboring hRBCs can be achieved using NOG-C3ΔMG2-3 mice, which could facilitate studies of human diseases associated with RBCs.

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In Vivo Efficacy of Bcl11a Erythroid-Enhancer Deletion in Humanized Mouse Models of Anemia

Blood ◽

10.1182/blood.v128.22.2465.2465 ◽

2016 ◽

Vol 128 (22) ◽

pp. 2465-2465 ◽

Cited By ~ 1

Author(s):

Thomas M Ryan ◽

Suean Daimia Fontenard ◽

Shanrun Liu ◽

Jonathan Lockhart ◽

Michael Berlett

Keyword(s):

Mouse Models ◽

Fetal Hemoglobin ◽

Thalassemia Major ◽

Beta Thalassemia ◽

Globin Genes ◽

Humanized Mouse ◽

Gamma Globin ◽

Hemoglobin Switching ◽

Humanized Mouse Models

Abstract Autologous cell therapy holds great promise for the treatment of beta thalassemia major and hemoglobinopathies like sickle cell anemia. Gene editing of a patient's own stem cells to reactivate the silenced gamma globin gene is one approach under active development. Prior to directly testing these new therapies in patients, we can answer some basic questions about their in vivo efficiency and efficacy in humanized mouse models of anemia. These models have their endogenous adult alpha and beta globin genes replaced with human alpha, gamma, and beta globin genes. These mice synthesize high level of human fetal hemoglobin during fetal life and complete their fetal-to-adult hemoglobin switch after birth. Experimental strategies designed to reactivate the silenced fetal gamma globin genes in adult erythroid cells are easily tested in vivo in these humanized hemoglobin switching mouse models. The silenced human fetal gamma globin genes can be activated by mutating the erythroid-specific enhancer of Bcl11a by gene editing. CRISPR sgRNAs, designed to target the +62 kb DNase I hypersensitive site in the second intron of Bcl11a, were microinjected along with Cas9 mRNA, into fertilized mouse embryos collected from humanized hemoglobin (Hb A) mice. The indel mutations that were generated in the founder animals were characterized and bred to homozygosity. The data demonstrates that the sgRNAs tested were successful in creating multiple unique mutations at the erythroid enhancer target sites. These mutations were transmitted through the germline allowing the effect of individual edited alleles to be analyzed. The majority of the mutations showed marginal increases in the number of F-cells over control animals. Significantly, despite having homozygous mutation of the erythroid-enhancer in all cells, fetal hemoglobin expression remains heterocellular. Importantly, the therapeutic efficacy of reactivating fetal hemoglobin with specific Bcl11a erythroid-enhancer mutations for the treatment of beta thalassemia major and sickle cell anemia was directly measured in vivo in these humanized models of disease. The reactivation of gamma globin in these humanized mouse models provides us with an opportunity to further interrogate the Bcl11a enhancer element, identify additional factors involved in hemoglobin switching and elucidate the mechanism driving pancellular vs heterocellular fetal hemoglobin expression. Disclosures No relevant conflicts of interest to declare.

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156 Humanized Mouse Models for The In Vivo Analysis of HIV Infection

JAIDS Journal of Acquired Immune Deficiency Syndromes ◽

10.1097/01.qai.0000351111.95546.8c ◽

2009 ◽

Vol 51 ◽

pp. 1

Author(s):

J Victor Garcia-Martinez

Keyword(s):

Hiv Infection ◽

Mouse Models ◽

Humanized Mouse ◽

In Vivo Analysis ◽

Humanized Mouse Models

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Humanized Mouse Models for the Advancement of Innate Lymphoid Cell-Based Cancer Immunotherapies

Frontiers in Immunology ◽

10.3389/fimmu.2021.648580 ◽

2021 ◽

Vol 12 ◽

Author(s):

Nina B. Horowitz ◽

Imran Mohammad ◽

Uriel Y. Moreno-Nieves ◽

Ievgen Koliesnik ◽

Quan Tran ◽

...

Keyword(s):

Nk Cells ◽

Mouse Models ◽

Human Cell Line ◽

Lymphoid Cells ◽

Humanized Mouse ◽

Cancer Treatments ◽

Human Genes ◽

Humanized Mouse Models ◽

And Behavior

Innate lymphoid cells (ILCs) are a branch of the immune system that consists of diverse circulating and tissue-resident cells, which carry out functions including homeostasis and antitumor immunity. The development and behavior of human natural killer (NK) cells and other ILCs in the context of cancer is still incompletely understood. Since NK cells and Group 1 and 2 ILCs are known to be important for mediating antitumor immune responses, a clearer understanding of these processes is critical for improving cancer treatments and understanding tumor immunology as a whole. Unfortunately, there are some major differences in ILC differentiation and effector function pathways between humans and mice. To this end, mice bearing patient-derived xenografts or human cell line-derived tumors alongside human genes or human immune cells represent an excellent tool for studying these pathways in vivo. Recent advancements in humanized mice enable unparalleled insights into complex tumor-ILC interactions. In this review, we discuss ILC behavior in the context of cancer, the humanized mouse models that are most commonly employed in cancer research and their optimization for studying ILCs, current approaches to manipulating human ILCs for antitumor activity, and the relative utility of various mouse models for the development and assessment of these ILC-related immunotherapies.

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Modelling of human herpesvirus infections in humanized mice

Journal of General Virology ◽

10.1099/vir.0.067793-0 ◽

2014 ◽

Vol 95 (10) ◽

pp. 2106-2117 ◽

Cited By ~ 7

Author(s):

Bradford K. Berges ◽

Anne Tanner

Keyword(s):

Mouse Models ◽

Immune Cells ◽

Human Herpesvirus ◽

Antiviral Drugs ◽

Humanized Mice ◽

Great Promise ◽

Humanized Mouse ◽

Herpes Simplex Viruses ◽

Human Immune ◽

Humanized Mouse Models

The human herpesviruses (HHVs) are remarkably successful human pathogens, with some members of the family successfully establishing infection in the vast majority of humans worldwide. Although many HHV infections result in asymptomatic infection or mild disease, there are rare cases of severe disease and death found with nearly every HHV. Many of the pathogenic mechanisms of these viruses are poorly understood, and in many cases, effective antiviral drugs are lacking. Only a single vaccine exists for the HHVs and researchers have been unable to develop treatments to cure the persistent infections associated with HHVs. A major hindrance to HHV research has been the lack of suitable animal models, with the notable exception of the herpes simplex viruses. One promising area for HHV research is the use of humanized mouse models, in which human cells or tissues are transplanted into immunodeficient mice. Current humanized mouse models mostly transplant human haematopoietic stem cells (HSCs), resulting in the production of a variety of human immune cells. Although all HHVs are thought to infect human immune cells, the beta- and gammaherpesviruses extensively infect and establish latency in these cells. Thus, mice humanized with HSCs hold great promise to study these herpesviruses. In this review, we provide a historical perspective on the use of both older and newer humanized mouse models to study HHV infections. The focus is on current developments in using humanized mice to study mechanisms of HHV-induced pathogenesis, human immune responses to HHVs and effectiveness of antiviral drugs.

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Peripheral blood CD34+ cells efficiently engraft human cytokine knock-in mice

Blood ◽

10.1182/blood-2015-10-676452 ◽

2016 ◽

Vol 128 (14) ◽

pp. 1829-1833 ◽

Cited By ~ 39

Author(s):

Yasuyuki Saito ◽

Jana M. Ellegast ◽

Anahita Rafiei ◽

Yuanbin Song ◽

Daniel Kull ◽

...

Keyword(s):

Immune System ◽

Mouse Models ◽

Humanized Mouse ◽

In Vivo Models ◽

Peripheral Blood Cd34 ◽

Cd34 Cells ◽

Key Points ◽

Humanized Mouse Models ◽

Human Cytokine

Key Points Human cytokine knock-in mice are improved in vivo models for multilineage engraftment of mobilized PB CD34+ cells. Humanized mouse models might open new avenues for personalized studies of human pathophysiology of the hematopoietic and immune system.

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Humanized mice

Journal of Experimental Medicine ◽

10.1084/jem.20051547 ◽

2005 ◽

Vol 202 (10) ◽

pp. 1307-1311 ◽

Cited By ~ 107

Author(s):

Francesca Macchiarini ◽

Markus G. Manz ◽

A. Karolina Palucka ◽

Leonard D. Shultz

Keyword(s):

Genetic Diversity ◽

Human Physiology ◽

Animal Models ◽

Infectious Diseases ◽

Mouse Models ◽

Humanized Mice ◽

Humanized Mouse ◽

Humanized Mouse Models ◽

The Creation ◽

Physiological Systems

Animal models have been instrumental in increasing the understanding of human physiology, particularly immunity. However, these animal models have been limited by practical considerations and genetic diversity. The creation of humanized mice that carry partial or complete human physiological systems may help overcome these obstacles. The National Institute of Allergy and Infectious Diseases convened a workshop on humanized mouse models for immunity in Bethesda, MD, on June 13–14, 2005, during which researchers discussed the benefits and limitations of existing animal models and offered insights into the development of future humanized mouse models.

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Translating Treg Therapy for Inflammatory Bowel Disease in Humanized Mice

Cells ◽

10.3390/cells10081847 ◽

2021 ◽

Vol 10 (8) ◽

pp. 1847

Author(s):

Sushmita Negi ◽

Sheetal Saini ◽

Nikunj Tandel ◽

Kiran Sahu ◽

Ravi P.N. Mishra ◽

...

Keyword(s):

Inflammatory Bowel Disease ◽

Immune Response ◽

T Cells ◽

Mouse Models ◽

Bowel Disease ◽

Humanized Mice ◽

Humanized Mouse ◽

Inflammatory Bowel ◽

Humanized Mouse Models

Crohn’s disease and ulcerative colitis, two major forms of inflammatory bowel disease (IBD) in humans, afflicted in genetically predisposed individuals due to dysregulated immune response directed against constituents of gut flora. The defective immune responses mounted against the regulatory mechanisms amplify and maintain the IBD-induced mucosal inflammation. Therefore, restoring the balance between inflammatory and anti-inflammatory immunepathways in the gut may contribute to halting the IBD-associated tissue-damaging immune response. Phenotypic and functional characterization of various immune-suppressive T cells (regulatory T cells; Tregs) over the last decade has been used to optimize the procedures for in vitro expansion of these cells for developing therapeutic interventional strategies. In this paper, we review the mechanisms of action and functional importance of Tregs during the pathogenesis of IBD and modulating the disease induced inflammation as well as role of mouse models including humanized mice repopulated with the human immune system (HIS) to study the IBD. “Humanized” mouse models provide new tools to analyze human Treg ontogeny, immunobiology, and therapy and the role of Tregs in developing interventional strategies against IBD. Overall, humanized mouse models replicate the human conditions and prove a viable tool to study molecular functions of human Tregs to harness their therapeutic potential.

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