Standardized assay for assessment of minimal residual disease in blood, bone marrow and apheresis from patients with plasma cell myeloma

Blum, Agnieszka; Haussmann, Katy; Streitz, Mathias; Schlickeiser, Stephan; Tietze-Buerger, Carola; Blau, Igor Wolfgang; Uharek, Lutz

doi:10.1038/s41598-019-39631-2

Cited by 11 publications

(9 citation statements)

References 37 publications

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“…The DURAclone technology used here has been successfully applied for the detection of regulatory T cells (Tregs), naïve/memory T cells and other immune cells in the peripheral blood of healthy human donors in previous work (33)(34)(35). Furthermore, the feasibility of this technology for immunomonitoring of cancer patients has been proven by the detection of minimal residual disease (MRD) in myeloma patients and by analyzing blood cells of cancer patients in response to immunotherapeutic nanoparticles (36,37). In addition, the introduced technology is not limited to CD19-targeting CAR approaches and can be readily adapted to the evaluation of other CAR immunotherapies.…”

Section: Discussionmentioning

confidence: 99%

Advanced Flow Cytometry Assays for Immune Monitoring of CAR-T Cell Applications

et al. 2021

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Adoptive immunotherapy using chimeric antigen receptor (CAR)-T cells has achieved successful remissions in refractory B-cell leukemia and B-cell lymphomas. In order to estimate both success and severe side effects of CAR-T cell therapies, longitudinal monitoring of the patient’s immune system including CAR-T cells is desirable to accompany clinical staging. To conduct research on the fate and immunological impact of infused CAR-T cells, we established standardized 13-colour/15-parameter flow cytometry assays that are suitable to characterize immune cell subpopulations in the peripheral blood during CAR-T cell treatment. The respective staining technology is based on pre-formulated dry antibody panels in a uniform format. Additionally, further antibodies of choice can be added to address specific clinical or research questions. We designed panels for the anti-CD19 CAR-T therapy and, as a proof of concept, we assessed a healthy individual and three B-cell lymphoma patients treated with anti-CD19 CAR-T cells. We analyzed the presence of anti-CD19 CAR-T cells as well as residual CD19+ B cells, the activation status of the T-cell compartment, the expression of co-stimulatory signaling molecules and cytotoxic agents such as perforin and granzyme B. In summary, this work introduces standardized and modular flow cytometry assays for CAR-T cell clinical research, which could also be adapted in the future as quality controls during the CAR-T cell manufacturing process.

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Section: Discussionmentioning

confidence: 99%

Advanced Flow Cytometry Assays for Immune Monitoring of CAR-T Cell Applications

et al. 2021

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“…For further characterization, the dose of multipotent progenitors among mobilized high stem cell can be phenotypically determined since multipotent progenitors might impact engraftment and immune reconstitution after stem cell transplantation 25 . Moreover, the use of an 8-color pre-formulated dried panel for the detection of minimal residual disease (MRD) was validated and its use can increase the quality of autologous blood stem cell products from myeloma patient apheresis to avoid the infusion of a residual disease 26 .…”

Section: Discussionmentioning

confidence: 99%

Widely applicable, extended flow cytometric stem cell enumeration panel for quality control of advanced cellular products

Haussmann

Streitz

Takvorian

et al. 2022

Sci Rep

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The most widely used quality control assay for CD34 + hematopoietic stem cell product characterization is the protocol established by the International Society of Hematotherapy and Graft Engineering (ISHAGE). While this protocol is still the gold standard for stem cell enumeration and viability assessment, it does not include T cell enumeration, which is nowadays mandatory for assaying standard allogeneic grafts and various advanced therapy medicinal products (ATMPs). In accordance, we have developed and extensively validated a new approach for a more comprehensive characterization of hematopoietic cellular products using a pre-formulated dried antibody format panel. In addition to the counting beads, the typical markers CD45 fluorescein isothiocyanate (FITC) and CD34 phycoerythrin (PE), as well as the viability dye 7-amino actinomycin D (7-AAD), our novel pre-formulated panel also contains CD3 Pacific Blue (PB) and CD19 allophycocyanin (APC) in the same tube, thereby allowing a combined calculation of leucocytes, stem cells, T and B cells. Showing high linearity, sensitivity and accuracy, our approach is easy to implement and enables a more in-depth characterization of the cellular product under release testing conditions. In addition, the dried pre-formulated antibody approach increases assay reliability compared to the standard antibody panel.

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“…To date, multiparameter flow cytometry (MFC; four or more colors) has been frequently used for the clinical detection of MRD [7][8][9][10][11][12][13][14][15]. However, more sensitive MFC methods for the assessment of MRD and prognostication of MM, including EuroFlow next-generation flow (EuroFlow-NGF) [16], MSKCC 10-color MFC [17], and DuraClone MFC [18], were recently developed and introduced. To accommodate these developments, the recent IMWG MRD criteria define a "flow MRD-negative" status as a CR plus MRDnegative status determined by EuroFlow-NGF or an equivalent method (sensitivity = 10 −5 ).…”

Section: Multiparameter Flow Cytometrymentioning

confidence: 99%

Clinical value of measurable residual disease testing for multiple myeloma and implementation in Japan

Takamatsu

2020

Int J Hematol

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The development of novel therapeutic agents has led to an increase in patients with multiple myeloma (MM) who achieve a complete response (CR). Consequently, a good correlation has been established between the CR rate and progression-free survival, and new methods are needed to stratify CR cases based on the measurable residual disease (MRD) and thus predict prognosis. Previously, multiparameter flow cytometry (MFC), which is rapid and widely available, has been used to assess MRD in patients with MM. Although the EuroFlow next-generation flow method was developed as a highly sensitive and standardized method of MRD detection, the procedure is costly within the current Japanese public medical insurance system. Recently, two Japanese clinical laboratory test companies, SRL and BML, respectively developed new and inexpensive 8-and 10-color single-tube MFC methods intended for the assessment of MRD under the current Japanese public medical insurance system. In this article, I have reviewed the most recent updates on MRD monitoring protocols in Japan and the clinical trials of the use of this parameter in patients with MM.

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Standardized assay for assessment of minimal residual disease in blood, bone marrow and apheresis from patients with plasma cell myeloma

Cited by 11 publications

References 37 publications

Advanced Flow Cytometry Assays for Immune Monitoring of CAR-T Cell Applications

Advanced Flow Cytometry Assays for Immune Monitoring of CAR-T Cell Applications

Widely applicable, extended flow cytometric stem cell enumeration panel for quality control of advanced cellular products

Clinical value of measurable residual disease testing for multiple myeloma and implementation in Japan

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