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Release of TcdA and TcdB from Clostridium difficile cdi 630 is not affected by functional inactivation of the tcdE gene

Olling, Alexandra ; Seehase, Sophie ; Minton, Nigel P. ; [et al.]

Elsevier BV ; 2012

In: Microbial Pathogenesis Vol. 52, No. 1 ( 2012-01), p. 92-100

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In: Microbial Pathogenesis, Elsevier BV, Vol. 52, No. 1 ( 2012-01), p. 92-100

Type of Medium: Online Resource

ISSN: 0882-4010

URL: Article

DOI: 10.1016/j.micpath.2011.10.009

Language: English

Publisher: Elsevier BV

Publication Date: 2012

detail.hit.zdb_id: 1471158-8

SSG: 12

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Endothelial protein C receptor supports hematopoietic stem cell engraftment and expansion in Mpl-deficient mice

Kohlscheen, Saskia ; Schenk, Franziska ; Rommel, Marcel G. E. ; [et al.]

American Society of Hematology ; 2019

In: Blood Vol. 133, No. 13 ( 2019-03-28), p. 1465-1478

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In: Blood, American Society of Hematology, Vol. 133, No. 13 ( 2019-03-28), p. 1465-1478

Abstract: Thrombopoietin (Thpo)/myeloproliferative leukemia virus oncogene (Mpl) signaling controls hematopoietic stem cell (HSC) self-renewal and quiescence; however, how these 2 seemingly opposing functions are controlled is not well understood. By transplantation of lentiviral-transduced hematopoietic cells in the Mpl-deficient mouse model, we addressed whether known or predicted Thpo target genes were able to rescue the Mpl-deficient phenotype of the mice. Among the tested genes, we identified endothelial protein C receptor (Epcr) to expand HSCs with the long-term (LT)-HSC surface phenotype in Mpl−/− mice and to enable secondary transplantation of Mpl-deficient bone marrow (BM). Epcr-transduced Mpl−/− HSCs enter quiescence earlier after transplantation than control-transduced Mpl−/− cells, and upregulated expression of the anti-apoptotic gene Bcl-xL. Also, in the wild-type background, Epcr expression marked the engrafting population in the BM. Furthermore, Epcr expression in Mpl−/− hematopoiesis increased the number of megakaryocytes in the BM. In vitro Thpo supported the surface expression of Epcr on primary murine hematopoietic stem and progenitor cells. With these data, we add new insights into Thpo-dependent influence on HSC engraftment after transplantation. This may be of use for the in vitro manipulation of HSCs, also in the context of gene therapy.

Type of Medium: Online Resource

ISSN: 0006-4971 , 1528-0020

URL: Article

DOI: 10.1182/blood-2018-03-837344

RVK:

XA 33000

RVK:

XA 10000

Language: English

Publisher: American Society of Hematology

Publication Date: 2019

detail.hit.zdb_id: 1468538-3

detail.hit.zdb_id: 80069-7

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Acute Influenza A Virus Infection Affects Cell Cycle and Lineage Output of Hematopoietic Stem Cells

Rommel, Marcel E G ; Walz, Lisa ; Kohlscheen, Saskia ; [et al.]

American Society of Hematology ; 2019

In: Blood Vol. 134, No. Supplement_1 ( 2019-11-13), p. 2467-2467

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In: Blood, American Society of Hematology, Vol. 134, No. Supplement_1 ( 2019-11-13), p. 2467-2467

Abstract: Long-term hematopoietic stem cells (LT-HSC) persist in quiescence to maintain their hematopoietic potential throughout life. In the case of need LT-HSC can be activated to replenish the pool of blood cells. We investigated the impact of acute influenza A virus (IAV) infection on hematopoiesis in C57Bl/6N mice, focusing on the most immature HSC and progenitors. Mice were infected with a lethal dose of IAV PR/8/1934 H1N1 (humane endpoints reached within 6 days post infection (dpi)). In two further groups, mice were treated daily with oseltamivir (antiviral neuraminidase inhibitor, dpi 0-4) or were vaccinated with single-cycle vesicular stomatitis virus replicon particles expressing a miss-matched neuraminidase from influenza A virus Yamaguchi/7/2004 H5N1 four weeks prior to infection. Both treatments rescued mice from infection-induced mortality. Every day 6-9 mice were analyzed for differences in the bone marrow (BM) and blood by flow cytometry and multiplex cytokine assays as well as in the lung to determine viral tissue titers and histopathology. HSC functionality was analyzed in a competitive BM transplantation of infected and non-infected mice. Irrespective of the treatment, high IAV lung tissue titers (≥5x106 tissue culture infectious dose 50) in the first days post infection (dpi 1-5) were associated with activation of HSC into the cell cycle. LT-HSCs (LSK, CD150+, CD34- and CD48-) were 50% less quiescent and shifted into the G1/S-G2-M phase (dpi 2-6) and returned to quiescence state after virus clearance (dpi 10). Furthermore, we detected 1.5-fold increase in proliferation of phenotypic LT-HSC. Differentiation was increased towards lymphoid progenitors (≥3-fold more compared to non-infected mice) during the acute phase of infection in untreated and oseltamivir treated mice and myeloid progenitors were reduced ~50% in all groups (dpi 4-8). We found the inflammatory cytokines IFNγ, IL-1α, IL-6, and TNFα to be significantly upregulated in the BM of untreated and oseltamivir treated mice but less in vaccinated animals (dpi 2-4). IL-1α or IL-6 stimulation of LT-HSCs was sufficient to initiate proliferation in cell culture. In all groups the initial drop of the peripheral platelet count (~30% lower compared to non-infected mice, dpi 2) was replenished with an excessive production of platelets (~45% increased, dpi 8-15). Histopathology and electron microscopy revealed the sequestration and accumulation of platelets in pulmonary capillaries and vessels. Subsequently, we detected twice as many mature megakaryocytes in the BM (dpi 2-4) and elevated CD41 expression on HSCs (LSK, CD150+ and CD34-; 3-fold more compared to non-infected mice dpi 2-6) indicating a myeloid/platelet-biased HSC compartment in response to infection. Competitive whole BM transplantation with activated LT-HSCs from the acute phase of infection vs non-infected mice showed delayed reconstitution of T-cells but a preferential differentiation towards platelets in recipient mice. Taken together, local IAV infection in the lung substantially affected LT-HSC quiescence and differentiation by inflammatory cytokines with systemic consequences and a myeloid/platelet-biased lineage output. Disclosures No relevant conflicts of interest to declare.

Type of Medium: Online Resource

ISSN: 0006-4971 , 1528-0020

URL: Article

DOI: 10.1182/blood-2019-126482

RVK:

XA 33000

RVK:

XA 10000

Language: English

Publisher: American Society of Hematology

Publication Date: 2019

detail.hit.zdb_id: 1468538-3

detail.hit.zdb_id: 80069-7

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Inhibition of Thrombopoietin/Mpl Signaling in Adult Hematopoiesis Identifies New Candidates for Hematopoietic Stem Cell Maintenance

Kohlscheen, Saskia ; Wintterle, Sabine ; Schwarzer, Adrian ; [et al.]

Public Library of Science (PLoS) ; 2015

In: PLOS ONE Vol. 10, No. 7 ( 2015-7-6), p. e0131866-

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In: PLOS ONE, Public Library of Science (PLoS), Vol. 10, No. 7 ( 2015-7-6), p. e0131866-

Type of Medium: Online Resource

ISSN: 1932-6203

URL: Article

DOI: 10.1371/journal.pone.0131866

DOI: 10.1371/journal.pone.0131866.g001

DOI: 10.1371/journal.pone.0131866.g002

DOI: 10.1371/journal.pone.0131866.g003

DOI: 10.1371/journal.pone.0131866.g004

DOI: 10.1371/journal.pone.0131866.g005

DOI: 10.1371/journal.pone.0131866.g006

DOI: 10.1371/journal.pone.0131866.g007

DOI: 10.1371/journal.pone.0131866.t001

DOI: 10.1371/journal.pone.0131866.s001

DOI: 10.1371/journal.pone.0131866.s002

DOI: 10.1371/journal.pone.0131866.s003

DOI: 10.1371/journal.pone.0131866.s004

DOI: 10.1371/journal.pone.0131866.s005

DOI: 10.1371/journal.pone.0131866.s006

DOI: 10.1371/journal.pone.0131866.s007

DOI: 10.1371/journal.pone.0131866.s008

DOI: 10.1371/journal.pone.0131866.s009

DOI: 10.1371/journal.pone.0131866.s010

DOI: 10.1371/journal.pone.0131866.s011

DOI: 10.1371/journal.pone.0131866.s012

DOI: 10.1371/journal.pone.0131866.s013

DOI: 10.1371/journal.pone.0131866.s014

DOI: 10.1371/journal.pone.0131866.s015

Language: English

Publisher: Public Library of Science (PLoS)

Publication Date: 2015

detail.hit.zdb_id: 2267670-3

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Signaling properties of murine MPL and MPL mutants after stimulation with thrombopoietin and romiplostim

Rommel, Marcel G.E. ; Hoerster, Keven ; Milde, Christian ; [et al.]

Elsevier BV ; 2020

In: Experimental Hematology Vol. 85 ( 2020-05), p. 33-46.e6

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In: Experimental Hematology, Elsevier BV, Vol. 85 ( 2020-05), p. 33-46.e6

Type of Medium: Online Resource

ISSN: 0301-472X

URL: Article

DOI: 10.1016/j.exphem.2020.04.006

RVK:

XA 42470

Language: English

Publisher: Elsevier BV

Publication Date: 2020

detail.hit.zdb_id: 2005403-8

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Promises and Challenges in Hematopoietic Stem Cell Gene Therapy

Kohlscheen, Saskia ; Bonig, Halvard ; Modlich, Ute

Mary Ann Liebert Inc ; 2017

In: Human Gene Therapy Vol. 28, No. 10 ( 2017-10), p. 782-799

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In: Human Gene Therapy, Mary Ann Liebert Inc, Vol. 28, No. 10 ( 2017-10), p. 782-799

Type of Medium: Online Resource

ISSN: 1043-0342 , 1557-7422

URL: Article

DOI: 10.1089/hum.2017.141

Language: English

Publisher: Mary Ann Liebert Inc

Publication Date: 2017

detail.hit.zdb_id: 1498064-2

SSG: 12

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3172 – VIRAL RESPIRATORY INFECTION INDUCES EMERGENCY MEGAKARYOPOIESIS AND GENERATES HYPER-REACTIVE PLATELETS BY A THROMBOPOIETIN INDEPENDENT MECHANISM

Rommel, Marcel GE ; Walz, Lisa ; Fotopoulou, Foteini ; [et al.]

Elsevier BV ; 2022

In: Experimental Hematology Vol. 111 ( 2022), p. S130-S131

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In: Experimental Hematology, Elsevier BV, Vol. 111 ( 2022), p. S130-S131

Type of Medium: Online Resource

ISSN: 0301-472X

URL: Article

DOI: 10.1016/j.exphem.2022.07.228

RVK:

XA 42470

Language: English

Publisher: Elsevier BV

Publication Date: 2022

detail.hit.zdb_id: 2005403-8

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Flow cytometric minimal residual disease assessment in B‐cell precursor acute lymphoblastic leukaemia patients treated with CD19‐targeted therapies — a EuroFlow study

Verbeek, Martijn W. C. ; Buracchi, Chiara ; Laqua, Anna ; [et al.]

Wiley ; 2022

In: British Journal of Haematology Vol. 197, No. 1 ( 2022-04), p. 76-81

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In: British Journal of Haematology, Wiley, Vol. 197, No. 1 ( 2022-04), p. 76-81

Abstract: The standardized EuroFlow protocol, including CD19 as primary B‐cell marker, enables highly sensitive and reliable minimal residual disease (MRD) assessment in B‐cell precursor acute lymphoblastic leukaemia (BCP‐ALL) patients treated with chemotherapy. We developed and validated an alternative gating strategy allowing reliable MRD analysis in BCP‐ALL patients treated with CD19‐targeting therapies. Concordant data were obtained in 92% of targeted therapy patients who remained CD19‐positive, whereas this was 81% in patients that became (partially) CD19‐negative. Nevertheless, in both groups median MRD values showed excellent correlation with the original MRD data, indicating that, despite higher interlaboratory variation, the overall MRD analysis was correct.

Type of Medium: Online Resource

ISSN: 0007-1048 , 1365-2141

URL: Issue

URL: Article

DOI: 10.1111/bjh.v197.1

DOI: 10.1111/bjh.17992

RVK:

XA 34100

Language: English

Publisher: Wiley

Publication Date: 2022

detail.hit.zdb_id: 1475751-5

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A ubiquitous chromatin opening element prevents transgene silencing in pluripotent stem cells and their differentiated progeny

Pfaff, Nils ; Lachmann, Nico ; Ackermann, Mania ; [et al.]

Oxford University Press (OUP) ; 2013

In: Stem Cells Vol. 31, No. 3 ( 2013-03-01), p. 488-499

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In: Stem Cells, Oxford University Press (OUP), Vol. 31, No. 3 ( 2013-03-01), p. 488-499

Abstract: Methylation-induced gene silencing represents a major obstacle to efficient transgene expression in pluripotent cells and thereof derived tissues. As ubiquitous chromatin opening elements (UCOE) have been shown to prevent transgene silencing in cell lines and primary hematopoietic cells, we hypothesized a similar activity in pluripotent cells. This concept was investigated in the context of cytidine deaminase (CDD) gene transfer, an approach to render hematopoietic cells resistant to the chemotherapeutic agent Ara-C. When murine induced pluripotent stem cells (iPSC)/embryonic stem cells (ESCs) were transduced with self-inactivating lentiviral vectors using housekeeping (truncated elongation factor 1α; EFS) or viral (spleen focus-forming virus; SFFV) promoters, incorporation of an heterogeneous nuclear ribonucleoproteins A2 B1/chromobox protein homolog 3 locus-derived UCOE (A2UCOE) significantly increased transgene expression and Ara-C resistance and effectively prevented silencing of the SFFV-promoter. The EFS promoter showed relatively stable transgene expression in naïve iPSCs, but rapid transgene silencing was observed upon hematopoietic differentiation. When combined with the A2UCOE, however, the EFS promoter yielded stable transgene expression in 73% ± 6% of CD41+ hematopoietic progeny, markedly increased CDD expression levels, and significantly enhanced Ara-C resistance in clonogenic cells. Bisulfite sequencing revealed protection from differentiation-induced promoter CpG methylation to be associated with these effects. Similar transgene promoting activities of the A2UCOE were observed during murine neurogenic differentiation, in naïve human pluripotent cells, and during nondirected multilineage differentiation of these cells. Thus, our data provide strong evidence that UCOEs can efficiently prevent transgene silencing in iPS/ESCs and their differentiated progeny and thereby introduce a generalized concept to circumvent differentiation-induced transgene silencing during the generation of advanced iPSC/ESC-based gene and cell therapy products.

Type of Medium: Online Resource

ISSN: 1066-5099 , 1549-4918

URL: Article

DOI: 10.1002/stem.1316

Language: English

Publisher: Oxford University Press (OUP)

Publication Date: 2013

detail.hit.zdb_id: 2030643-X

detail.hit.zdb_id: 1143556-2

detail.hit.zdb_id: 605570-9

SSG: 12

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Efficient Hematopoietic Redifferentiation of Induced Pluripotent Stem Cells Derived from Primitive Murine Bone Marrow Cells

Pfaff, Nils ; Lachmann, Nico ; Kohlscheen, Saskia ; [et al.]

Mary Ann Liebert Inc ; 2012

In: Stem Cells and Development Vol. 21, No. 5 ( 2012-03-20), p. 689-701

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In: Stem Cells and Development, Mary Ann Liebert Inc, Vol. 21, No. 5 ( 2012-03-20), p. 689-701

Type of Medium: Online Resource

ISSN: 1547-3287 , 1557-8534

URL: Article

DOI: 10.1089/scd.2011.0010

Language: English

Publisher: Mary Ann Liebert Inc

Publication Date: 2012

detail.hit.zdb_id: 2142305-2

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