TY  - JOUR
A1  - Koeniger, Tobias
A1  - Bell, Luisa
A1  - Mifka, Anika
A1  - Enders, Michael
A1  - Hautmann, Valentin
A1  - Mekala, Subba Rao
A1  - Kirchner, Philipp
A1  - Ekici, Arif B.
A1  - Schulz, Christian
A1  - Wörsdörfer, Philipp
A1  - Mencl, Stine
A1  - Kleinschnitz, Christoph
A1  - Ergün, Süleyman
A1  - Kuerten, Stefanie
T1  - Bone marrow‐derived myeloid progenitors in the leptomeninges of adult mice
JF  - Stem Cells
N2  - Although the bone marrow contains most hematopoietic activity during adulthood, hematopoietic stem and progenitor cells can be recovered from various extramedullary sites. Cells with hematopoietic progenitor properties have even been reported in the adult brain under steady‐state conditions, but their nature and localization remain insufficiently defined. Here, we describe a heterogeneous population of myeloid progenitors in the leptomeninges of adult C57BL/6 mice. This cell pool included common myeloid, granulocyte/macrophage, and megakaryocyte/erythrocyte progenitors. Accordingly, it gave rise to all major myelo‐erythroid lineages in clonogenic culture assays. Brain‐associated progenitors persisted after tissue perfusion and were partially inaccessible to intravenous antibodies, suggesting their localization behind continuous blood vessel endothelium such as the blood‐arachnoid barrier. Flt3\(^{Cre}\) lineage tracing and bone marrow transplantation showed that the precursors were derived from adult hematopoietic stem cells and were most likely continuously replaced via cell trafficking. Importantly, their occurrence was tied to the immunologic state of the central nervous system (CNS) and was diminished in the context of neuroinflammation and ischemic stroke. Our findings confirm the presence of myeloid progenitors at the meningeal border of the brain and lay the foundation to unravel their possible functions in CNS surveillance and local immune cell production.
KW  - hematopoietic
KW  - meninges
KW  - mouse
KW  - myeloid
KW  - progenitor
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-224452
VL  - 39
IS  - 2
SP  - 227
EP  - 239
ER  - 
TY  - JOUR
A1  - Majumder, Snigdha
A1  - Jugovic, Isabelle
A1  - Saul, Domenica
A1  - Bell, Luisa
A1  - Hundhausen, Nadine
A1  - Seal, Rishav
A1  - Beilhack, Andreas
A1  - Rosenwald, Andreas
A1  - Mougiakakos, Dimitrios
A1  - Berberich-Siebelt, Friederike
T1  - Rapid and Efficient Gene Editing for Direct Transplantation of Naive Murine Cas9\(^+\) T Cells
JF  - Frontiers in Immunology
N2  - Gene editing of primary T cells is a difficult task. However, it is important for research and especially for clinical T-cell transfers. CRISPR/Cas9 is the most powerful gene-editing technique. It has to be applied to cells by either retroviral transduction or electroporation of ribonucleoprotein complexes. Only the latter is possible with resting T cells. Here, we make use of Cas9 transgenic mice and demonstrate nucleofection of pre-stimulated and, importantly, of naive CD3\(^+\) T cells with guideRNA only. This proved to be rapid and efficient with no need of further selection. In the mixture of Cas9\(^+\)CD3\(^+\) T cells, CD4\(^+\) and CD8\(^+\) conventional as well as regulatory T cells were targeted concurrently. IL-7 supported survival and naivety in vitro, but T cells were also transplantable immediately after nucleofection and elicited their function like unprocessed T cells. Accordingly, metabolic reprogramming reached normal levels within days. In a major mismatch model of GvHD, not only ablation of NFATc1 and/or NFATc2, but also of the NFAT-target gene IRF4 in naïve primary murine Cas9\(^+\)CD3\(^+\) T cells by gRNA-only nucleofection ameliorated GvHD. However, pre-activated murine T cells could not achieve long-term protection from GvHD upon single NFATc1 or NFATc2 knockout. This emphasizes the necessity of gene-editing and transferring unstimulated human T cells during allogenic hematopoietic stem cell transplantation.
KW  - CRISPR/Cas9
KW  - gRNA-only
KW  - GvHD
KW  - metabolism
KW  - NFAT
KW  - naive T-cell gene editing
KW  - T-cell transfer
KW  - IRF4
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-242896
SN  - 1664-3224
VL  - 12
ER  -