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Article ; Online: A 19-color single-tube full spectrum flow cytometry assay for the detection of measurable residual disease in acute myeloid leukemia.

Fokken, Hendrik / Waclawski, Julian / Kattre, Nadine / Kloos, Arnold / Müller, Sebastian / Ettinger, Max / Kacprowski, Tim / Heuser, Michael / Maetzig, Tobias / Schwarzer, Adrian

Cytometry. Part A : the journal of the International Society for Analytical Cytology

2023 Volume 105, Issue 3, Page(s) 181–195

Abstract: Multiparameter flow cytometry (MFC) has emerged as a standard method for quantifying measurable residual disease (MRD) in acute myeloid leukemia. However, the limited number of available channels on conventional flow cytometers requires the division of a ...

Abstract	Multiparameter flow cytometry (MFC) has emerged as a standard method for quantifying measurable residual disease (MRD) in acute myeloid leukemia. However, the limited number of available channels on conventional flow cytometers requires the division of a diagnostic sample into several tubes, restricting the number of cells and the complexity of immunophenotypes that can be analyzed. Full spectrum flow cytometers overcome this limitation by enabling the simultaneous use of up to 40 fluorescent markers. Here, we used this approach to develop a good laboratory practice-conform single-tube 19-color MRD detection assay that complies with recommendations of the European LeukemiaNet Flow-MRD Working Party. We based our assay on clinically-validated antibody clones and evaluated its performance on an IVD-certified full spectrum flow cytometer. We measured MRD and normal bone marrow samples and compared the MRD data to a widely used reference MRD-MFC panel generating highly concordant results. Using our newly developed single-tube panel, we established reference values in healthy bone marrow for 28 consensus leukemia-associated immunophenotypes and introduced a semi-automated dimensionality-reduction, clustering and cell type identification approach that aids the unbiased detection of aberrant cells. In summary, we provide a comprehensive full spectrum MRD-MFC workflow with the potential for rapid implementation for routine diagnostics due to reduced cell requirements and ease of data analysis with increased reproducibility in comparison to conventional FlowMRD routines.
MeSH term(s)	Humans ; Flow Cytometry/methods ; Reproducibility of Results ; Leukemia, Myeloid, Acute/diagnosis ; Bone Marrow/metabolism ; Neoplasm, Residual/diagnosis
Language	English
Publishing date	2023-12-08
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2099868-5
ISSN	1552-4930 ; 0196-4763 ; 1552-4922
ISSN (online)	1552-4930
ISSN	0196-4763 ; 1552-4922
DOI	10.1002/cyto.a.24811
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Book ; Online ; Thesis: Modifikation des Tropismus onkolytischer Adenoviren mittels bispezifischer Adapterproteine für die Therapie Polysialinsäure-exprimierender Tumoren

Kloos, Arnold [Verfasser]

2013

Author's details	Arnold Kloos
Keywords	Biowissenschaften, Biologie ; Life Science, Biology
Subject code	sg570
Language	German
Publisher	Technische Informationsbibliothek und Universitätsbibliothek Hannover (TIB)
Publishing place	Hannover
Document type	Book ; Online ; Thesis
Database	Digital theses on the web

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Article ; Online: Fludarabine, cytarabine, and idarubicin with or without venetoclax in patients with relapsed/refractory acute myeloid leukemia.

Haematologica

2024 Volume 109, Issue 1, Page(s) 72–83

Abstract: Treatment options for relapsed and refractory acute myeloid leukemia patients (R/R AML) are limited. This retrospective cohort study compares safety and efficacy of fludarabine, cytarabine, and idarubicin (FLA-IDA) without or with venetoclax (FLAVIDA) in ...

Abstract	Treatment options for relapsed and refractory acute myeloid leukemia patients (R/R AML) are limited. This retrospective cohort study compares safety and efficacy of fludarabine, cytarabine, and idarubicin (FLA-IDA) without or with venetoclax (FLAVIDA) in patients with R/R AML. Thirty-seven and 81 patients received one course FLA-IDA with or without a 7-day course of venetoclax, respectively. The overall response rate (ORR) was significantly higher in FLAVIDA compared to FLAIDA- treated patients (78% vs. 47%; P=0.001), while measurable residual disease was negative at a similar proportion in responding patients (50% vs. 57%), respectively. Eighty-one percent and 79% of patients proceeded to allogeneic hematopoietic cell transplantation or donor lymphocyte infusion after FLAVIDA and FLA-IDA, respectively. Event-free and overall survival were similar in FLAVIDA- and FLA-IDA-treated patients. Refractory patients could be salvaged more successfully after FLA-IDA compared to FLAVIDA pretreatment. Neutrophil and platelet recovery times were similar in the venetoclax and the control group. In conclusion, short-term venetoclax in combination with FLA-IDA represents an effective treatment regimen in R/R AML identifying chemosensitive patients rapidly and inducing measurable residual disease-negative remission in a high proportion of R/R AML patients.
MeSH term(s)	Humans ; Idarubicin/therapeutic use ; Cytarabine ; Retrospective Studies ; Granulocyte Colony-Stimulating Factor ; Leukemia, Myeloid, Acute/drug therapy ; Vidarabine ; Antineoplastic Combined Chemotherapy Protocols/adverse effects
Chemical Substances	Idarubicin (ZRP63D75JW) ; Cytarabine (04079A1RDZ) ; venetoclax (N54AIC43PW) ; fludarabine (P2K93U8740) ; Granulocyte Colony-Stimulating Factor (143011-72-7) ; Vidarabine (FA2DM6879K)
Language	English
Publishing date	2024-01-01
Publishing country	Italy
Document type	Journal Article
ZDB-ID	2333-4
ISSN	1592-8721 ; 0017-6567 ; 0390-6078
ISSN (online)	1592-8721
ISSN	0017-6567 ; 0390-6078
DOI	10.3324/haematol.2023.282912
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Article: Improved Activity against Acute Myeloid Leukemia with Chimeric Antigen Receptor (CAR)-NK-92 Cells Designed to Target CD123

Morgan, Michael A. / Kloos, Arnold / Lenz, Daniela / Kattre, Nadine / Nowak, Juliette / Bentele, Marco / Keisker, Maximilian / Dahlke, Julia / Zimmermann, Katharina / Sauer, Martin / Heuser, Michael / Schambach, Axel

Viruses. 2021 July 14, v. 13, no. 7

2021

Abstract: Anti-cancer activity can be improved by engineering immune cells to express chimeric antigen receptors (CARs) that recognize tumor-associated antigens. Retroviral vector gene transfer strategies allow stable and durable transgene expression. Here, we ... ...

Abstract	Anti-cancer activity can be improved by engineering immune cells to express chimeric antigen receptors (CARs) that recognize tumor-associated antigens. Retroviral vector gene transfer strategies allow stable and durable transgene expression. Here, we used alpharetroviral vectors to modify NK-92 cells, a natural killer cell line, with a third-generation CAR designed to target the IL-3 receptor subunit alpha (CD123), which is strongly expressed on the surface of acute myeloid leukemia (AML) cells. Alpharetroviral vectors also contained a transgene cassette to allow constitutive expression of human IL-15 for increased NK cell persistence in vivo. The anti-AML activity of CAR-NK-92 cells was tested via in vitro cytotoxicity assays with the CD123⁺ AML cell line KG-1a and in vivo in a patient-derived xenotransplantation CD123⁺ AML model. Unmodified NK-92 cells or NK-92 cells modified with a truncated version of the CAR that lacked the signaling domain served as controls. Alpharetroviral vector-modified NK-92 cells stably expressed the transgenes and secreted IL-15. Anti-CD123-CAR-NK-92 cells exhibited enhanced anti-AML activity in vitro and in vivo as compared to control NK-92 cells. Our data (1) shows the importance of IL-15 expression for in vivo persistence of NK-92 cells, (2) supports continued investigation of anti-CD123-CAR-NK cells to target AML, and (3) points towards potential strategies to further improve CAR-NK anti-AML activity.
Keywords	antigens ; antineoplastic activity ; cell lines ; cytotoxicity ; gene expression ; gene transfer ; humans ; interleukin-15 ; interleukin-3 ; models ; myeloid leukemia ; natural killer cells ; retroviral vectors ; transgenes ; xenotransplantation
Language	English
Dates of publication	2021-0714
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2516098-9
ISSN	1999-4915
ISSN	1999-4915
DOI	10.3390/v13071365
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Scalable generation of functional human iPSC-derived CAR-macrophages that efficiently eradicate CD19-positive leukemia.

Abdin, Shifaa M / Paasch, Daniela / Kloos, Arnold / Oliveira, Marco Carvalho / Jang, Mi-Sun / Ackermann, Mania / Stamopoulou, Andriana / Mroch, Philipp J / Falk, Christine S / von Kaisenberg, Constantin S / Schambach, Axel / Heuser, Michael / Moritz, Thomas / Hansen, Gesine / Morgan, Michael / Lachmann, Nico

Journal for immunotherapy of cancer

2023 Volume 11, Issue 12

Abstract: Background: Macrophages have recently become attractive therapeutics in cancer immunotherapy. The potential of macrophages to infiltrate and influence solid malignancies makes them promising targets for the chimeric antigen receptor (CAR) technology to ... ...

Abstract	Background: Macrophages have recently become attractive therapeutics in cancer immunotherapy. The potential of macrophages to infiltrate and influence solid malignancies makes them promising targets for the chimeric antigen receptor (CAR) technology to redirect their stage of polarization, thus enhancing their anticancer capacities. Given the emerging interest for CAR-macrophages, generation of such cells so far mainly depends on peripheral blood monocytes, which are isolated from the respective donor prior to genetic manipulation. This procedure is time-intensive and cost-intensive, while, in some cases, insufficient monocyte amounts can be recovered from the donor, thus hampering the broad applicability of this technology. Hence, we demonstrate the generation and effectiveness of CAR-macrophages from various stem cell sources using also modern upscaling technologies for next generation immune cell farming. Methods: Primary human hematopoietic stem and progenitor cells and induced pluripotent stem cells were used to derive anti-CD19 CAR-macrophages. Anticancer activity of the cells was demonstrated in co-culture systems, including primary material from patients with leukemia. Generation of CAR-macrophages was facilitated by bioreactor technologies and single-cell RNA (scRNA) sequencing was used to characterize in-depth response and behavior of CAR-macrophages. Results: Irrespective of the stem-cell source, CAR-macrophages exhibited enhanced and antigen-dependent phagocytosis of CD19 Conclusion: In summary, our work provides insights into the seminal use and behavior of CAR-macrophages which are derived from various sources of stem cells, while introducing a unique technology for CAR-macrophage manufacturing, all dedicated to the clinical translation of CAR-macrophages within the field of anticancer immunotherapies.
MeSH term(s)	Humans ; Receptors, Chimeric Antigen/genetics ; Receptors, Antigen, T-Cell ; Induced Pluripotent Stem Cells/metabolism ; T-Lymphocytes ; Leukemia/therapy ; Neoplasms ; Macrophages/metabolism
Chemical Substances	Receptors, Chimeric Antigen ; Receptors, Antigen, T-Cell
Language	English
Publishing date	2023-12-22
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2719863-7
ISSN	2051-1426 ; 2051-1426
ISSN (online)	2051-1426
ISSN	2051-1426
DOI	10.1136/jitc-2023-007705
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Article ; Online: Improved Activity against Acute Myeloid Leukemia with Chimeric Antigen Receptor (CAR)-NK-92 Cells Designed to Target CD123.

Morgan, Michael A / Kloos, Arnold / Lenz, Daniela / Kattre, Nadine / Nowak, Juliette / Bentele, Marco / Keisker, Maximilian / Dahlke, Julia / Zimmermann, Katharina / Sauer, Martin / Heuser, Michael / Schambach, Axel

Viruses

2021 Volume 13, Issue 7

Abstract	Anti-cancer activity can be improved by engineering immune cells to express chimeric antigen receptors (CARs) that recognize tumor-associated antigens. Retroviral vector gene transfer strategies allow stable and durable transgene expression. Here, we used alpharetroviral vectors to modify NK-92 cells, a natural killer cell line, with a third-generation CAR designed to target the IL-3 receptor subunit alpha (CD123), which is strongly expressed on the surface of acute myeloid leukemia (AML) cells. Alpharetroviral vectors also contained a transgene cassette to allow constitutive expression of human IL-15 for increased NK cell persistence in vivo. The anti-AML activity of CAR-NK-92 cells was tested via in vitro cytotoxicity assays with the CD123
MeSH term(s)	Aged ; Alpharetrovirus/genetics ; Animals ; Cell Line, Tumor ; Female ; Genetic Therapy ; Genetic Vectors/genetics ; Humans ; Immunotherapy, Adoptive/methods ; Interleukin-3 Receptor alpha Subunit/genetics ; Interleukin-3 Receptor alpha Subunit/immunology ; Interleukin-3 Receptor alpha Subunit/metabolism ; Killer Cells, Natural/immunology ; Leukemia, Myeloid, Acute/drug therapy ; Leukemia, Myeloid, Acute/genetics ; Leukemia, Myeloid, Acute/immunology ; Male ; Mice ; Mice, Inbred NOD ; Primary Cell Culture ; Receptors, Chimeric Antigen/genetics ; Receptors, Chimeric Antigen/immunology ; Transduction, Genetic ; Transgenes ; Xenograft Model Antitumor Assays
Chemical Substances	IL3RA protein, human ; Interleukin-3 Receptor alpha Subunit ; Receptors, Chimeric Antigen
Language	English
Publishing date	2021-07-14
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v13071365
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Article: Retargeted oncolytic viruses provoke tumor-directed T-cell responses.

Kloos, Arnold / Woller, Norman / Gerardy-Schahn, Rita / Kühnel, Florian

Oncoimmunology

2015 Volume 4, Issue 12, Page(s) e1052933

Abstract: Intratumoral application of oncolytic viruses effectively induce tumor-directed immune responses. However, their systemic application is typically insufficient to stimulate the required extent of tumor tissue inflammation to elicit antitumor immunity. We ...

Abstract	Intratumoral application of oncolytic viruses effectively induce tumor-directed immune responses. However, their systemic application is typically insufficient to stimulate the required extent of tumor tissue inflammation to elicit antitumor immunity. We recently discovered evidence that this barrier can be overcome by effective molecular retargeting of viral infection.
Language	English
Publishing date	2015-08-20
Publishing country	United States
Document type	Journal Article
ZDB-ID	2645309-5
ISSN	2162-402X ; 2162-4011
ISSN (online)	2162-402X
ISSN	2162-4011
DOI	10.1080/2162402X.2015.1052933
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Article ; Online: Correction: Effective drug treatment identified by in vivo screening in a transplantable patient-derived xenograft model of chronic myelomonocytic leukemia.

Kloos, Arnold / Mintzas, Konstantinos / Winckler, Lina / Gabdoulline, Razif / Alwie, Yasmine / Jyotsana, Nidhi / Kattre, Nadine / Schottmann, Renate / Scherr, Michaela / Gupta, Charu / Adams, Felix F / Schwarzer, Adrian / Heckl, Dirk / Schambach, Axel / Imren, Suzan / Humphries, R Keith / Ganser, Arnold / Thol, Felicitas / Heuser, Michael

Leukemia

2021 Volume 35, Issue 12, Page(s) 3629

Language	English
Publishing date	2021-10-30
Publishing country	England
Document type	Published Erratum
ZDB-ID	807030-1
ISSN	1476-5551 ; 0887-6924
ISSN (online)	1476-5551
ISSN	0887-6924
DOI	10.1038/s41375-021-01459-z
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Article ; Online: Clinical Implications and Dynamics of Clonal Hematopoiesis in Anti-CD19 CAR T-cell Treated Patients.

Panagiota, Victoria / Kerschbaum, Johanna Franziska / Penack, Olaf / Stein, Catarina M / Arends, Christopher M / Koenecke, Christian / Strzelecka, Paulina M / Kloos, Arnold / Wiegand, Laura / Lasch, Alina / Altwasser, Robert / Halik, Adriane / Gabdoulline, Razif / Thomson, Julia / Weibl, Konstantin / Franke, Georg-Nikolaus / Berger, Carolina / Hasenkamp, Justin / Ayuk, Francis /

Na, Il-Kang / Beutel, Gernot / Keller, Ulrich / Bullinger, Lars / Wulf, Gerald Georg / Kröger, Nicolaus / Vucinic, Vladan / Heuser, Michael / Damm, Frederik

HemaSphere

2023 Volume 7, Issue 10, Page(s) e957

Abstract: Recent evidence revealed important interactions between clonal hematopoiesis (CH) and cellular therapies established for the treatment of hematologic malignancies. The impact of CH on safety, efficacy, and outcome of chimeric antigen receptor (CAR) T- ... ...

Abstract	Recent evidence revealed important interactions between clonal hematopoiesis (CH) and cellular therapies established for the treatment of hematologic malignancies. The impact of CH on safety, efficacy, and outcome of chimeric antigen receptor (CAR) T-cell therapy is currently under investigation. We analyzed 110 patients with relapsed/refractory B-cell non-Hodgkin lymphoma (n = 105) or acute lymphoblastic leukemia (ALL) (n = 5), treated with Axicabtagene-Ciloleucel (39%), Tisagenlecleucel (51%), or Brexucabtagene autoleucel (10%). Using error-corrected targeted sequencing, a high CH prevalence of 56.4% (variant allele frequency [VAF] ≥1%) at the time of CAR T-cell infusion was detected. The most frequently mutated gene was
Language	English
Publishing date	2023-10-03
Publishing country	United States
Document type	Journal Article
ISSN	2572-9241
ISSN (online)	2572-9241
DOI	10.1097/HS9.0000000000000957
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Article ; Online: Molecular retargeting of antibodies converts immune defense against oncolytic viruses into cancer immunotherapy.

Niemann, Julia / Woller, Norman / Brooks, Jennifer / Fleischmann-Mundt, Bettina / Martin, Nikolas T / Kloos, Arnold / Knocke, Sarah / Ernst, Amanda M / Manns, Michael P / Kubicka, Stefan / Wirth, Thomas C / Gerardy-Schahn, Rita / Kühnel, Florian

Nature communications

2019 Volume 10, Issue 1, Page(s) 3236

Abstract: Virus-neutralizing antibodies are a severe obstacle in oncolytic virotherapy. Here, we present a strategy to convert this unfavorable immune response into an anticancer immunotherapy via molecular retargeting. Application of a bifunctional adapter ... ...

Abstract	Virus-neutralizing antibodies are a severe obstacle in oncolytic virotherapy. Here, we present a strategy to convert this unfavorable immune response into an anticancer immunotherapy via molecular retargeting. Application of a bifunctional adapter harboring a tumor-specific ligand and the adenovirus hexon domain DE1 for engaging antiadenoviral antibodies, attenuates tumor growth and prolongs survival in adenovirus-immunized mice. The therapeutic benefit achieved by tumor retargeting of antiviral antibodies is largely due to NK cell-mediated triggering of tumor-directed CD8 T-cells. We further demonstrate that antibody-retargeting (Ab-retargeting) is a feasible method to sensitize tumors to PD-1 immune checkpoint blockade. In therapeutic settings, Ab-retargeting greatly improves the outcome of intratumor application of an oncolytic adenovirus and facilitates long-term survival in treated animals when combined with PD-1 checkpoint inhibition. Tumor-directed retargeting of preexisting or virotherapy-induced antiviral antibodies therefore represents a promising strategy to fully exploit the immunotherapeutic potential of oncolytic virotherapy and checkpoint inhibition.
MeSH term(s)	Adenoviridae/genetics ; Adenoviridae/immunology ; Animals ; Antibodies/immunology ; Antibodies, Neutralizing/immunology ; CD8-Positive T-Lymphocytes/immunology ; Cell Line, Tumor ; HEK293 Cells ; Humans ; Immunotherapy/methods ; Killer Cells, Natural/immunology ; Mice ; Molecular Targeted Therapy/methods ; Neoplasms/immunology ; Neoplasms/therapy ; Oncolytic Virotherapy/methods ; Oncolytic Viruses/genetics ; Oncolytic Viruses/immunology
Chemical Substances	Antibodies ; Antibodies, Neutralizing
Language	English
Publishing date	2019-07-19
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-019-11137-5
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