A novel model to study neonatalEscherichia colisepsis and the effect of treatment on the human immune system using humanized mice

AbstractBackgroundHumanized mice featuring a functional human immune system are an important pre-clinical model for examining immune responses to human-specific pathogens. This model has been widely utilized to study human diseases that are otherwise impossible or difficult to investigate in humans or with other animal models. However, one limitation of using humanized mice is their native murine gut microbiome, which significantly differs from the one found in humans. These differences may be even greater for mice housed and bred in specific pathogen free conditions. Given the importance of the gut microbiome to human health and disease, these differences may profoundly impact the ability to translate the results from humanized mice studies to human disease. Further, there is a critical need for improved pre-clinical models to study the complex in vivo relationships of the gut microbiome, immune system, and human disease. We therefore created double humanized mice with both a functional human immune system and stable human-like gut microbiome.ResultsSurgery was performed on NOD.Cg-PrkdcscidII2rgtm1Wjl/SzJ (NSG) mice to create bone-marrow, liver, thymus (BLT) humanized mice. After immune reconstitution, mice were treated with broad spectrum antibiotics to deplete murine gut bacteria and then transplanted with fecal material from healthy human donors. Characterization of 173 fecal samples obtained from 45 humanized mice revealed that double humanized mice had unique 16S rRNA gene profiles consistent with those of the individual human donor samples. Importantly, transplanted human-like gut microbiomes were stable in mice for the duration of the study, up to 14.5 weeks post-transplant. Microbiomes of double humanized mice also harbored predicted functional capacities that more closely resembled those of the human donors compared to humanized mice.ConclusionsHere, we describe successful engraftment of a stable human microbiome in BLT humanized mice to further improve this preclinical humanized mouse model. These double humanized mice represent a unique and tractable new model to study the complex relationships between the human gut microbiome, human immune system, and human disease in vivo.

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Human immune cells infiltrate the lesioned spinal cord and impair recovery after spinal cord injury in humanized mice

10.1101/626721 ◽

2019 ◽

Cited By ~ 1

Author(s):

Randall S. Carpenter ◽

Roselyn R. Jiang ◽

Faith H. Brennan ◽

Jodie C.E. Hall ◽

Manoj K. Gottipati ◽

...

Keyword(s):

Spinal Cord ◽

Immune System ◽

Humanized Mice ◽

Human Umbilical Cord ◽

Cns Injury ◽

Human Immune System ◽

Neuroinflammatory Response ◽

Human T Cells ◽

Cord Injury ◽

Human Immune

Summary PointsImmune compromised mice require ~4 months of engraftment with human umbilical cord blood CD34+ stem cells to develop a full and functional human immune systemThe human neuroinflammatory response elicited after spinal cord injury in humanized mice is limited at 2 months post-engraftment but matures by 4 monthsIntraspinal neuroinflammation consists of a florid human T cell and macrophage response, and human T cells co-localize with human macrophagesA human intraspinal neuroinflammatory response exacerbates lesion pathology and impairs functional recoveryAbstractHumanized mice are a useful tool to help better understand how the human immune system responds to central nervous system (CNS) injury. However, the optimal parameters for using humanized mice in preclinical CNS injury models have not been established. Here, we show that it takes 3-4 months after engraftment of neonatal immune compromised mice with human umbilical cord stem cells to generate a robust human immune system. Indeed, sub-optimal human immune cell responses occurred when humanized mice received spinal contusion injuries at 2 months vs. 4 months post-engraftment. Human T cells directly contact human macrophages within the spinal cord lesion of these mice and the development of a mature human immune system was associated with worse lesion pathology and neurological recovery. Together, data in this report establish an optimal experimental framework for using humanized mice to help translate promising preclinical therapies for CNS injury.

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Building the Next Generation of Humanized Hemato-Lymphoid System Mice

Frontiers in Immunology ◽

10.3389/fimmu.2021.643852 ◽

2021 ◽

Vol 12 ◽

Author(s):

Tijana Martinov ◽

Kelly M. McKenna ◽

Wei Hong Tan ◽

Emily J. Collins ◽

Allie R. Kehret ◽

...

Keyword(s):

Immune System ◽

Adaptive Immunity ◽

Humanized Mice ◽

Human Immune System ◽

Functional Interactions ◽

Life Saving ◽

Mouse Tissues ◽

Fundamental Biology ◽

Human Immune

Since the late 1980s, mice have been repopulated with human hematopoietic cells to study the fundamental biology of human hematopoiesis and immunity, as well as a broad range of human diseases in vivo. Multiple mouse recipient strains have been developed and protocols optimized to efficiently generate these “humanized” mice. Here, we review three guiding principles that have been applied to the development of the currently available models: (1) establishing tolerance of the mouse host for the human graft; (2) opening hematopoietic niches so that they can be occupied by human cells; and (3) providing necessary support for human hematopoiesis. We then discuss four remaining challenges: (1) human hematopoietic lineages that poorly develop in mice; (2) limited antigen-specific adaptive immunity; (3) absent tolerance of the human immune system for its mouse host; and (4) sub-functional interactions between human immune effectors and target mouse tissues. While major advances are still needed, the current models can already be used to answer specific, clinically-relevant questions and hopefully inform the development of new, life-saving therapies.

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Antigen-specific human T-cell responses and T cell–dependent production of human antibodies in a humanized mouse model

Blood ◽

10.1182/blood-2007-11-121319 ◽

2008 ◽

Vol 111 (8) ◽

pp. 4293-4296 ◽

Cited By ~ 85

Author(s):

Noriko Tonomura ◽

Katsuyoshi Habiro ◽

Akira Shimizu ◽

Megan Sykes ◽

Yong-Guang Yang

Keyword(s):

Immune System ◽

T Cell ◽

Humanized Mice ◽

Human Immune System ◽

Hematopoietic Stem ◽

Fetal Thymus ◽

Human Antibodies ◽

Human Immune ◽

Liver Tissues

Abstract Humanized mice with a functional human immune system would be very useful for in vivo studies of human immunobiology. We have previously shown that cotransplantation of human fetal thymus/liver tissues and CD34+ fetal liver cells into immunodeficient nonobese diabetic severe combined immunodeficiency (NOD/SCID) mice leads to the development of multiple lineages of human lymphohematopoietic cells and formation of secondary lymphoid organs with normal architecture. Here, we evaluated the ability of these humanized mice to develop antigen-specific, T cell–dependent antibody responses after in vivo immunization with T-dependent antigen, 2,4-dinitrophenyl hapten-keyhole limpet hemocyanin (DNP23-KLH). Human T cells from DNP23-KLH–immunized mice showed strong proliferation in response to KLH in vitro. Furthermore, T cell–dependent production of DNP-specific human antibodies (mainly IgG1 and IgG2) was detected in all immunized mice. These results confirm that a functional human immune system can be established in immunodeficient mice through cotransplantation of human fetal thymus/liver tissues and CD34+ hematopoietic stem/progenitor cells.

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Infection and immune control of human oncogenic γ-herpesviruses in humanized mice

Philosophical Transactions of the Royal Society B Biological Sciences ◽

10.1098/rstb.2018.0296 ◽

2019 ◽

Vol 374 (1773) ◽

pp. 20180296 ◽

Cited By ~ 11

Author(s):

Donal McHugh ◽

Nicole Caduff ◽

Anita Murer ◽

Christine Engelmann ◽

Yun Deng ◽

...

Keyword(s):

Immune System ◽

Kaposi Sarcoma ◽

Humanized Mice ◽

Cytotoxic Lymphocytes ◽

Chronic Infections ◽

Human Immune System ◽

Immune Control ◽

Barr Virus ◽

Human Immune

Epstein–Barr virus (EBV) and Kaposi sarcoma-associated herpesvirus (KSHV) comprise the oncogenic human γ-herpesvirus family and are responsible for 2–3% of all tumours in man. With their prominent growth-transforming abilities and high prevalence in the human population, these pathogens have probably shaped the human immune system throughout evolution for near perfect immune control of the respective chronic infections in the vast majority of healthy pathogen carriers. The exclusive tropism of EBV and KSHV for humans has, however, made it difficult in the past to study their infection, tumourigenesis and immune control in vivo . Mice with reconstituted human immune system components (humanized mice) support replication of both viruses with both persisting latent and productive lytic infection. Moreover, B-cell lymphomas can be induced by EBV alone and KSHV co-infection with gene expression hallmarks of human malignancies that are associated with both viruses. Furthermore, cell-mediated immune control by primarily cytotoxic lymphocytes is induced upon infection and can be probed for its functional characteristics as well as putative requirements for its priming. Insights that have been gained from this model and remaining questions will be discussed in this review. This article is part of the theme issue ‘Silent cancer agents: multi-disciplinary modelling of human DNA oncoviruses’.

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Potential Pitfalls of the Humanized Mice in Modeling Sepsis

International Journal of Inflammation ◽

10.1155/2018/6563454 ◽

2018 ◽

Vol 2018 ◽

pp. 1-9 ◽

Cited By ~ 5

Author(s):

Krzysztof Laudanski ◽

Michael Stentz ◽

Matthew DiMeglio ◽

William Furey ◽

Toby Steinberg ◽

...

Keyword(s):

Immune System ◽

Data Interpretation ◽

Humanized Mice ◽

Limiting Factors ◽

Human Immune System ◽

Human Immunology ◽

General Terms ◽

Protozoan Infections ◽

Human Immune ◽

Bone Marrow Mesenchymal Cells

Humanized mice are a state-of-the-art tool used to study several diseases, helping to close the gap between mice and human immunology. This review focuses on the potential obstacles in the analysis of immune system performance between humans and humanized mice in the context of severe acute inflammation as seen in sepsis or other critical care illnesses. The extent to which the reconstituted human immune system in mice adequately compares to the performance of the human immune system in human hosts is still an evolving question. Although certain viral and protozoan infections can be replicated in humanized mice, whether a highly complex and dynamic systemic inflammation like sepsis can be accurately represented by current humanized mouse models in a clinically translatable manner is unclear. Humanized mice are xenotransplant animals in the most general terms. Several organs (e.g., bone marrow mesenchymal cells, endothelium) cannot interact with the grafted human leukocytes effectively due to species specificity. Also the interaction between mice gut flora and the human immune system may be paradoxical. Often, grafting is performed utilizing an identical batch of stem cells in highly inbred animals which fails to account for human heterogeneity. Limiting factors include the substantial cost and restricting supply of animals. Finally, humanized mice offer an opportunity to gain knowledge of human-like conditions, requiring careful data interpretation just as in nonhumanized animals.

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Human immune cells infiltrate the spinal cord and impair recovery after spinal cord injury in humanized mice

Scientific Reports ◽

10.1038/s41598-019-55729-z ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 2

Author(s):

Randall S. Carpenter ◽

Roselyn R. Jiang ◽

Faith H. Brennan ◽

Jodie C. E. Hall ◽

Manoj K. Gottipati ◽

...

Keyword(s):

Spinal Cord ◽

Spinal Cord Injury ◽

Immune System ◽

Humanized Mice ◽

Human Umbilical Cord Blood ◽

Human Umbilical Cord ◽

Cns Injury ◽

Human Immune System ◽

Cord Injury ◽

Human Immune

AbstractHumanized mice can be used to better understand how the human immune system responds to central nervous system (CNS) injury and inflammation. The optimal parameters for using humanized mice in preclinical CNS injury models need to be established for appropriate use and interpretation. Here, we show that the developmental age of the human immune system significantly affects anatomical and functional outcome measures in a preclinical model of traumatic spinal cord injury (SCI). Specifically, it takes approximately 3–4 months for a stable and functionally competent human immune system to develop in neonatal immune compromised mice after they are engrafted with human umbilical cord blood stem cells. Humanized mice receiving a SCI before or after stable engraftment exhibit significantly different neuroinflammatory profiles. Importantly, the development of a mature human immune system was associated with worse lesion pathology and neurological recovery after SCI. In these mice, human T cells infiltrate the spinal cord lesion and directly contact human macrophages. Together, data in this report establish an optimal experimental framework for using humanized mice to help translate promising preclinical therapies for CNS injury.

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Humanized Mice, a New Model To Study the Influence of Drug Treatment on Neonatal Sepsis

Infection and Immunity ◽

10.1128/iai.01235-12 ◽

2013 ◽

Vol 81 (5) ◽

pp. 1520-1531 ◽

Cited By ~ 22

Author(s):

Wolfgang Ernst ◽

Nicole Zimara ◽

Frank Hanses ◽

Daniela N. Männel ◽

Birgit Seelbach-Göbel ◽

...

Keyword(s):

Immune Response ◽

Immune System ◽

Bacterial Infection ◽

Perinatal Care ◽

Bacterial Load ◽

Humanized Mice ◽

Western World ◽

Human Immune System ◽

New Model ◽

Human Immune

ABSTRACTBacterial infection with group BStreptococcus(GBS) represents a prominent threat to neonates and fetuses in the Western world, causing severe organ damage and even death. To improve current therapeutic strategies and to investigate new approaches, an appropriatein vivomodel to study the immune response of a human immune system is needed. Therefore, we introduced humanized mice as a new model for GBS-induced sepsis. Humanized mice feature deficiencies similar to those found in neonates, such as lower immunoglobulin levels and myeloid cell dysfunction. Due to the husbandry in specific-pathogen-free (SPF) facilities, the human immune cells in these mice also exhibit a naive phenotype which mimics the conditions in fetuses/neonates. Following infection, cytokine release and leukocyte trafficking from the bone marrow to the lymphoid organ (spleen) and into the peritoneum (site of infection) as well as bacterial spreading and clearance were traceable in the humanized mice. Furthermore, we investigated the effects of betamethasone and indomethacin treatment using this novel sepsis model. Although both drugs are commonly used in perinatal care, little is known about their effects on the neonatal immune system. Treatment of infected humanized mice not only induced the reduction of human leukocytes in the spleen but also increased the bacterial load in all analyzed organs, including the brain, which did not show infiltration of live GBS in untreated controls. These studies demonstrate the utility of the humanized mice as a new model to study an immature human immune response during bacterial infection and allow the investigation of side effects induced by various treatments.

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Characterization of human cancer xenografts in humanized mice

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2019-000416 ◽

2020 ◽

Vol 8 (1) ◽

pp. e000416 ◽

Cited By ~ 4

Author(s):

Jonathan Rios-Doria ◽

Christina Stevens ◽

Christopher Maddage ◽

Kerri Lasky ◽

Holly K Koblish

Keyword(s):

Immune System ◽

Immune Cell ◽

Human Cancer ◽

Humanized Mice ◽

Tumor Growth Rate ◽

Human Immune System ◽

Tumor Models ◽

Antitumor Response ◽

Human Immune

BackgroundPreclinical evaluation of drugs targeting the human immune system has posed challenges for oncology researchers. Since the commercial introduction of humanized mice, antitumor efficacy and pharmacodynamic studies can now be performed with human cancer cells within mice bearing components of a human immune system. However, development and characterization of these models is necessary to understand which model may be best suited for different agents.MethodsWe characterized A375, A549, Caki-1, H1299, H1975, HCC827, HCT116, KU-19–19, MDA-MB-231, and RKO human cancer cell xenografts in CD34+humanized non-obese diabetic-scid gamma mice for tumor growth rate, immune cell profiling, programmed death ligand 1 (PD-L1) expression and response to anti-PD-L1 therapy. Immune cell profiling was performed using flow cytometry and immunohistochemistry. Antitumor response of humanized xenograft models to PD-L1 therapy was performed using atezolizumab.ResultsWe found that CD4+and CD8+T-cell composition in both the spleen and tumor varied among models, with A375, Caki-1, MDA-MB-231, and HCC827 containing higher intratumoral frequencies of CD4+and CD8+T cells of CD45+cells compared with other models. We demonstrate that levels of immune cell infiltrate within each model are strongly influenced by the tumor and not the stem cell donor. Many of the tumor models showed an abundance of myeloid cells, B cells and dendritic cells. RKO and MDA-MB-231 tumors contained the highest expression of PD-L1+tumor cells. The antitumor response of the models to atezolizumab was positively associated with the level of CD4+and CD8+tumor-infiltrating lymphocytes (TILs).ConclusionsThese data demonstrate that there are tumor-intrinsic factors that influence the immune cell repertoire within tumors and spleen, and that TIL frequencies are a key factor in determining response to anti-PD-L1 in tumor xenografts in humanized mice. These data may also aid in the selection of tumor models to test antitumor activity of novel immuno-oncology or tumor-directed agents.

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A novel combination of chemotherapy and immunotherapy controls tumor growth in mice with a human immune system

10.1101/191031 ◽

2017 ◽

Author(s):

Aude Burlion ◽

Rodrigo N. Ramos ◽

KC Pukar ◽

Kélhia Sendeyo ◽

Aurélien Corneau ◽

...

Keyword(s):

Breast Cancer ◽

T Cells ◽

Dendritic Cells ◽

Immune System ◽

Regulatory T Cells ◽

Tumor Growth ◽

Humanized Mice ◽

Tumor Control ◽

Human Immune System ◽

Human Immune

AbstractMice reconstituted with a human immune system and bearing human tumors represent a promising model for developing novel cancer immunotherapies. Here, we used mass cytometry and multi-parametric flow cytometry to characterize human leukocytes infiltrating a human breast cancer tumor model in immunocompromised NOD.SCID.γc-null mice reconstituted with a human immune system and compared it to samples of breast cancer patients. We observed highly activated human CD4+ and CD8+ T cells in the tumor, as well as minor subsets of innate immune cells in both settings. We also report that ICOS+ CD4+ regulatory T cells (Treg) were enriched in the tumor relative to the periphery in humanized mice and patients, providing a target to affect Treg and tumor growth. Indeed, administration of a neutralizing mAb to human ICOS reduced Treg proportions and numbers and improved CD4+ T cell proliferation in humanized mice. Moreover, a combination of the anti-ICOS mAb with cyclophosphamide reduced tumor growth, and that was associated with an improved CD8 to Treg ratio. However, depletion of human CD8+ T cells only marginally affected tumor control whereas depletion of murine myeloid cells abrogated the effect of the combination therapy. Altogether, our results indicate that a combination of anti-ICOS mAb and chemotherapy controls tumor growth in humanized mice and highlight the crucial implication of innate immunity in treatment efficacy, opening new perspectives for the treatment of breast cancer.One sentence summaryICOS expressed on Tregs is a promising target to improve tumor immunity in humansAbbreviationsICOSInducible CostimulatoryNSGNOD.SCID.gc-nullTregregulatory T cellsCTXcyclophosphamideHuMicehumanized miceCyTOFcytometry time-of-flighttSNEtdistributed stochastic neighbor embeddingpDCsplasmacytoid dendritic cellsDCdendritic cellsICDimmunogenic cell death

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