Engraftment of human HSCs in nonirradiated newborn NOD-scid IL2r null mice is enhanced by transgenic expression of membrane-bound human SCF

Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605, USA.
Blood (Impact Factor: 10.45). 01/2012; 119(12):2778-88. DOI: 10.1182/blood-2011-05-353243
Source: PubMed

ABSTRACT Immunodeficient mice engrafted with human HSCs support multidisciplinary translational experimentation, including the study of human hematopoiesis. Heightened levels of human HSC engraftment are observed in immunodeficient mice expressing mutations in the IL2-receptor common γ chain (IL2rg) gene, including NOD-scid IL2rγ(null) (NSG) mice. Engraftment of human HSC requires preconditioning of immunodeficient recipients, usually with irradiation. Such preconditioning increases the expression of stem cell factor (SCF), which is critical for HSC engraftment, proliferation, and survival. We hypothesized that transgenic expression of human membrane-bound stem cell factor Tg(hu-mSCF)] would increase levels of human HSC engraftment in nonirradiated NSG mice and eliminate complications associated with irradiation. Surprisingly, detectable levels of human CD45(+) cell chimerism were observed after transplantation of cord blood-derived human HSCs into nonirradiated adult as well as newborn NSG mice. However, transgenic expression of human mSCF enabled heightened levels of human hematopoietic cell chimerism in the absence of irradiation. Moreover, nonirradiated NSG-Tg(hu-mSCF) mice engrafted as newborns with human HSCs rejected human skin grafts from a histoincompatible donor, indicating the development of a functional human immune system. These data provide a new immunodeficient mouse model that does not require irradiation preconditioning for human HSC engraftment and immune system development.

22 Reads
  • Source
    • "Such modifications might include replacing growth factors or matrix molecules that poorly cross-react between mouse and human species. A number of recently described strains, when crossed with NBSGW, might further improve the differentiation of human HSCs (Billerbeck et al., 2011; Brehm et al., 2012; Rongvaux et al., 2014). "
    [Show abstract] [Hide abstract]
    ABSTRACT: In this study, we demonstrate a newly derived mouse model that supports engraftment of human hematopoietic stem cells (HSCs) in the absence of irradiation. We cross the NOD.Cg-Prkdc(scid)Il2rg(tm1Wjl)/SzJ (NSG) strain with the C57BL/6J-Kit(W-41J)/J (C57BL/6.Kit(W41)) strain and engraft, without irradiation, the resulting NBSGW strain with human cord blood CD34+ cells. At 12-weeks postengraftment in NBSGW mice, we observe human cell chimerism in marrow (97% ± 0.4%), peripheral blood (61% ± 2%), and spleen (94% ± 2%) at levels observed with irradiation in NSG mice. We also detected a significant number of glycophorin-A-positive expressing cells in the developing NBSGW marrow. Further, the observed levels of human hematopoietic chimerism mimic those reported for both irradiated NSG and NSG-transgenic strains. This mouse model permits HSC engraftment while avoiding the complicating hematopoietic, gastrointestinal, and neurological side effects associated with irradiation and allows investigators without access to radiation to pursue engraftment studies with human HSCs. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.
    Stem Cell Reports 01/2015; 117(2). DOI:10.1016/j.stemcr.2014.12.005 · 5.37 Impact Factor
  • Source
    • "However, in BRgWv mice we found that SCF protein levels and transcripts in bone marrow niche cells are unaltered, suggesting that the increased HSC engraftment in these mice is independent of SCF expression/availability. In line with this hypothesis, prior work has shown that increased levels of human SCF can support increased differentiation, but not engraftment of human HSCs (Brehm et al., 2012 "
    [Show abstract] [Hide abstract]
    ABSTRACT: In-depth analysis of the cellular and molecular mechanisms regulating human HSC function will require a surrogate host that supports robust maintenance of transplanted human HSCs in vivo, but the currently available options are problematic. Previously we showed that mutations in the Kit receptor enhance engraftment of transplanted HSCs in the mouse. To generate an improved model for human HSC transplantation and analysis, we developed immune-deficient mouse strains containing Kit mutations. We found that mutation of the Kit receptor enables robust, uniform, sustained, and serially transplantable engraftment of human HSCs in adult mice without a requirement for irradiation conditioning. Using this model, we also showed that differential KIT expression identifies two functionally distinct subpopulations of human HSCs. Thus, we have found that the capacity of this Kit mutation to open up stem cell niches across species barriers has significant potential and broad applicability in human HSC research.
    Cell Stem Cell 07/2014; 15(2). DOI:10.1016/j.stem.2014.06.001 · 22.27 Impact Factor
  • Source
    • "The recent development of ‘humanized’ mice has allowed the in vivo investigation of human immune responses [17]–[20]. In transplantation models, we and others have demonstrated that ex vivo expanded human Tregs can prevent the development of transplant arteriosclerosis [21] and skin allograft rejections [22] in peripheral blood mononuclear cells (PBMC)-reconstituted humanized mouse models [23]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Type 1 diabetes mellitus (T1DM) is an autoimmune disease caused by immune-mediated destruction of insulin-secreting β cells of the pancreas. Near complete dependence on exogenous insulin makes T1DM very difficult to control, with the result that patients are exposed to high blood glucose and risk of diabetic complications and/or intermittent low blood glucose that can cause unconsciousness, fits and even death. Allograft transplantation of pancreatic islets restores normoglycemia with a low risk of surgical complications. However, although successful immediately after transplantation, islets are progressively lost, with most of the patients requiring exogenous insulin within 2 years post-transplant. Therefore, there is an urgent requirement for the development of new strategies to prevent islet rejection. In this study, we explored the importance of human regulatory T cells in the control of islets allograft rejection. We developed a pre-clinical model of human islet transplantation by reconstituting NOD-scid IL2rγnull mice with cord blood-derived human CD34+ stem cells and demonstrated that although the engrafted human immune system mediated the rejection of human islets, their survival was significantly prolonged following adoptive transfer of ex vivo expanded human Tregs. Mechanistically, Tregs inhibited the infiltration of innate immune cells and CD4+ T cells into the graft by down-regulating the islet graft-derived monocyte chemoattractant protein-1. Our findings might contribute to the development of clinical strategies for Treg therapy to control human islet rejection. We also show for the first time that CD34+ cells-reconstituted NOD-scid IL2rγnull mouse model could be beneficial for investigating human innate immunity in vivo.
    PLoS ONE 03/2014; 9(3):e90387. DOI:10.1371/journal.pone.0090387 · 3.23 Impact Factor
Show more