Mouse Models for Human Hemato‐Lymphopoiesis

Ziegler, Patrick; Manz, Markus G.

doi:10.1002/0471140856.tx0213s31

Cited by 3 publications

(1 citation statement)

References 15 publications

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“…After Ficoll gradient centrifugation CD34 þ cells were purified using a Midi-MACS CD34 isolation kit (Miltenyi Biotec, Bergisch-Gladbach, Germany), as previously described. 44 Purity of CD34 þ cells as determined by flow cytometry was above 95%.…”

Section: Purification Of Hematopoietic Progenitor Cells From Patient mentioning

confidence: 97%

Src family kinases mediate cytoplasmic retention of activated STAT5 in BCR–ABL-positive cells

et al. 2012

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Persistent activation of the Abl tyrosine kinase in the BCR-ABL fusion protein is the major cause of chronic myeloid leukemia (CML). Among many other substrates BCR-ABL phosphorylates STAT5 and Src family kinases (SFK). Activated pSTAT5 is essential for initial transformation and maintenance of the disease. Cytokine-induced phosphorylation on tyrosine 694 typically leads to nuclear accumulation of pSTAT5 and target gene expression. We verified that in BCR-ABL-positive progenitor cells from a CML patient and in K562 cells pSTAT5 is cytoplasmic. However, upon ectopic expression of BCR-ABL p210 in non-myeloid cells, co-transfected STAT5A is phosphorylated on Y694 and localized in the nucleus arguing for an additional factor mediating cytoplasmic retention in CML cells. Expression of the SFK v-Src, Hck or Lyn together with STAT5A results in phosphorylation on Y694 and cytoplasmic retention. Upon coexpression of BCR-ABL and individual SFK the cytoplasmic retention of activated STAT5A mediated by v-Src and Hck but not Lyn is dominant over nuclear translocation induced by BCR-ABL. Cytoplasmic retention depends on the kinase activity of SFK and is mediated through the interaction of the SH2 domain of STAT5A with the SFK. Interestingly, nuclear accumulation of STAT5A as a result of activation by FLT3-ITD, an oncogene found in acute myeloid leukemia, cannot be prevented by coexpression of SFK. Importantly, inhibition of SFK in K562 cells restored nuclear accumulation of pSTAT5A, enhanced STAT5 target gene expression and increased colony formation. Thus, SFK mediate cytoplasmic retention of pSTAT5A in BCR-ABL-positive cells. Cytoplasmic pSTAT5A in CML cells might balance the controversial functions of STAT5 in cellular senescence and differentiation versus G1/S progression and survival.

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Section: Purification Of Hematopoietic Progenitor Cells From Patient mentioning

confidence: 97%

Src family kinases mediate cytoplasmic retention of activated STAT5 in BCR–ABL-positive cells

et al. 2012

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Generation of Humanized Mice for Analysis of Human Dendritic Cells

Saito

Ellegast

Manz

2016

Methods in Molecular Biology

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Transplantation of human CD34(+) hematopoietic stem and progenitor cells into severe immunocompromised newborn mice allows the development of a human hemato-lymphoid system (HHLS) including dendritic cells (DCs) in vivo. Therefore, it can be a powerful tool to study human DC subsets, residing in different lymphoid and nonlymphoid organs. We have recently generated novel mouse strains called human cytokine knock-in mice in which human versions of several cytokines are knocked into Rag2(-/-)γC(-/-) strains. In addition, human SIRPα, which is a critical factor to prevent donor cell to be eliminated by host macrophages, is expressed as transgene. These mice efficiently support human myeloid cell development and, indeed, allow the analysis of three major subsets of human DC lineages, plasmacytoid DCs and CD1c(+) and CD141(+) classical DCs. Moreover, these strains also support cytokine-mobilized peripheral blood CD34(+) cell engraftment and subsequent DC development. Here we describe our standard methods to characterize DCs developed in human cytokine knock-in mice.

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Recognising the potential of large animals for modelling neuromuscular junction physiology and disease

et al. 2022

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The aetiology and pathophysiology of many diseases of the motor unit remain poorly understood and the role of the neuromuscular junction (NMJ) in this group of disorders is particularly overlooked, especially in humans, when these diseases are comparatively rare.However, elucidating the development, function and degeneration of the NMJ is essential to uncover its contribution to neuromuscular disorders, and to explore potential therapeutic avenues to treat these devastating diseases. Until now, an understanding of the role of the NMJ in disease pathogenesis has been hindered by inherent differences between rodent and human NMJs: stark contrasts in body size and corresponding differences in associated axon length underpin some of the translational issues in animal models of neuromuscular disease.Comparative studies in large mammalian models, including examination of naturallyoccurring, highly prevalent animal diseases and evaluating their treatment, might provide more relevant insight into the pathogenesis and therapy of equivalent human diseases. This review argues that large animal models offer great potential to enhance our understanding of the neuromuscular system in health and disease, and in particular when dealing with diseases for which nerve length dependency might underly the pathogenesis.

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Mouse Models for Human Hemato‐Lymphopoiesis

Cited by 3 publications

References 15 publications

Src family kinases mediate cytoplasmic retention of activated STAT5 in BCR–ABL-positive cells

Src family kinases mediate cytoplasmic retention of activated STAT5 in BCR–ABL-positive cells

Generation of Humanized Mice for Analysis of Human Dendritic Cells

Recognising the potential of large animals for modelling neuromuscular junction physiology and disease

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