| 1. |
- Johansson, Pegah, 1978, et al.
(författare)
-
Validation of a flow cytometry-based detection of γ-H2AX, to measure DNA damage for clinical applications.
- 2017
-
Ingår i: Cytometry. Part B, Clinical cytometry. - : Wiley. - 1552-4957 .- 1552-4949. ; 92, s. 534-540
-
Tidskriftsartikel (refereegranskat)abstract
- The nucleosomal histone protein H2AX is specifically phosphorylated (γ-H2AX) adjacent to DNA double-strand breaks (DSBs) and is used for quantifying DSBs. Many chemotherapies and ionizing radiation (IR) used in cancer treatment result in DSBs. Therefore, γ-H2AX has a significant potential as a biomarker in evaluating patient sensitivity and responsiveness to IR and chemotherapy.
|
|
| 2. |
|
|
| 3. |
- Langenskiöld, Cecilia, et al.
(författare)
-
Determination of blood cell subtype concentrations from frozen whole blood samples using TruCount beads
- 2018
-
Ingår i: Cytometry Part B - Clinical Cytometry. - : Wiley. - 1552-4949 .- 1552-4957. ; 94:4, s. 660-666
-
Tidskriftsartikel (refereegranskat)abstract
- © 2016 International Clinical Cytometry Society.Background: In many studies it would be advantageous if blood samples could be collected and analyzed using flow cytometry at a later stage. Ideally, sample collection should involve little hands-on time, allow for long-term storage, and minimally influence the samples. Methods: Here we establish a flow cytometry antibody panel that can be used to determine granulocytes, monocytes, and lymphocyte subset concentrations in fresh and frozen whole blood using TruCount technology. Results: The panel can be used on fresh whole-blood samples as well as whole-blood samples that have been frozen after mixing with 10% DMSO. Concentrations in frozen and fresh sample is highly correlated both when frozen within 4 h and the day after collection (r≥0.98), and the estimated concentration in frozen samples was between 91 and 94% of that in fresh samples for all cell types. Conclusion: Using this method whole-blood samples can be frozen using a simple preparation method, and stored long-term before accurate determination of cell concentration. This allows for standardized analysis of the samples at a reference laboratory in multi-center studies.
|
|
| 4. |
|
|
| 5. |
- Porwit, Anna, et al.
(författare)
-
Multiparameter Flow Cytometry Applications in the Diagnosis of Mixed Phenotype Acute Leukemia
- 2019
-
Ingår i: Cytometry Part B - Clinical Cytometry. - : Wiley. - 1552-4949 .- 1552-4957. ; 96:3, s. 183-194
-
Forskningsöversikt (refereegranskat)abstract
- Mixed phenotype acute leukemias (MPALs) represent a rare subgroup of acute leukemias with a poor prognosis. Proper diagnosis and classification of MPAL is extremely important for patients' outcome. Morphology and flow cytometry recognize two types of MPAL: the “bilineal” MPAL with the coexistence of two blast populations of different lineage and truly “biphenotypic” MPAL coexpressing markers of more than one lineage in a homogenous blast population, respectively. The WHO 2008 classification further delineated three categories: associated with t(9;22)/BCR-ABL1 fusion gene, associated with KMT2A (mixed lineage leukemia) rearrangements, and nonotherwise specified. These categories remained unchanged in the WHO2016 update. Molecular studies have further underlined the heterogeneity of MPAL. In this review, rules for the correct assignment of acute leukemia to the MPAL category are discussed, including both flow cytometry and immunohistochemistry on bone marrow or other tissues biopsies. Comparison of the immunophenotypic classification proposals is provided outlining the explorations mandatory for definitive diagnosis. An extensive review of published data summarizes the reported cytogenetic and molecular anomalies. New developments in the understanding of the early stages of hematopoiesis provide clues to the possible etiopathology of these diseases. Finally, current treatment recommendations are summarized and referenced for clinical use, pointing out that allogeneic hematopoietic stem cell transplantation at an early stage should be considered (at least in adult patients).
|
|
| 6. |
- Rawstron, Andy C., et al.
(författare)
-
Reproducible diagnosis of chronic lymphocytic leukemia by flow cytometry : An European Research Initiative on CLL (ERIC) & European Society for Clinical Cell Analysis (ESCCA) Harmonisation project
- 2018
-
Ingår i: Cytometry. Part B, Clinical cytometry.. - : Wiley. - 1552-4949 .- 1552-4957. ; 94:1, s. 121-128
-
Tidskriftsartikel (refereegranskat)abstract
- The diagnostic criteria for CLL rely on morphology and immunophenotype. Current approaches have limitations affecting reproducibility and there is no consensus on the role of new markers. The aim of this project was to identify reproducible criteria and consensus on markers recommended for the diagnosis of CLL. ERIC/ESCCA members classified 14 of 35 potential markers as “required” or “recommended” for CLL diagnosis, consensus being defined as >75% and >50% agreement, respectively. An approach to validate “required” markers using normal peripheral blood was developed. Responses were received from 150 participants with a diagnostic workload >20 CLL cases per week in 23/150 (15%), 5–20 in 82/150 (55%), and <5 cases per week in 45/150 (30%). The consensus for “required” diagnostic markers included: CD19, CD5, CD20, CD23, Kappa, and Lambda. “Recommended” markers potentially useful for differential diagnosis were: CD43, CD79b, CD81, CD200, CD10, and ROR1. Reproducible criteria for component reagents were assessed retrospectively in 14,643 cases from 13 different centers and showed >97% concordance with current approaches. A pilot study to validate staining quality was completed in 11 centers. Markers considered as “required” for the diagnosis of CLL by the participants in this study (CD19, CD5, CD20, CD23, Kappa, and Lambda) are consistent with current diagnostic criteria and practice. Importantly, a reproducible approach to validate and apply these markers in individual laboratories has been identified. Finally, a consensus “recommended” panel of markers to refine diagnosis in borderline cases (CD43, CD79b, CD81, CD200, CD10, and ROR1) has been defined and will be prospectively evaluated.
|
|
| 7. |
- Viktorisson, Adam, et al.
(författare)
-
A control for the day-to-day normalization of the flow cytometry γ-H2AX assay for clinical routine.
- 2018
-
Ingår i: Cytometry. Part B, Clinical cytometry. - : Wiley. - 1552-4957 .- 1552-4949. ; 94:6, s. 946-949
-
Tidskriftsartikel (refereegranskat)abstract
- The phosphorylation of histone H2AX (γ-H2AX) at the DNA double-strand break (DSB) site is frequently used for quantifying DSBs and may be useful as a biomarker for clinical applications. We have previously reported a flow cytometry-based quantification of γ-H2AX for clinical routine. One major challenge, however, is the lack of a control sample for normalization of the day-to-day variation of the flow cytometry γ-H2AX assay.Here, we report development of a mix-control sample containing peripheral blood mononuclear cells (PBMC) from 10 control individuals, for normalization of day-to-day variation of the flow cytometry-γ-H2AX assay.We showed that control individuals sampled on different days show an average day-to-day variation (CV) of 34%, which was reduced to 12% after normalization to the control sample. The normalization allowed detection of radiosensitivity of lymphoblastoid cell lines from ataxia telangiectasia patients, sampled over three days.The mix-control sample, consisting of 10 control individuals' PBMC, can be used as a control sample to normalize for day-to-day variation of the γ-H2AX assay. The use of this sample will facilitate integration of the γ-H2AX assay into clinical routine. © 2018 International Clinical Cytometry Society.
|
|
| 8. |
- Jafari, Katayoon, et al.
(författare)
-
Visualization of cell composition and maturation in the bone marrow using 10-color flow cytometry and radar plots
- 2018
-
Ingår i: Cytometry Part B - Clinical Cytometry. - : Wiley. - 1552-4949. ; 94:2, s. 219-229
-
Tidskriftsartikel (refereegranskat)abstract
- Background: The enormous potential of complex data files generated by 10-color flow cytometry (FC) is hindered by the requirement for exhaustive manual gating and the complexity of multidimensional data visualization. We propose a model using radar plots (RPs), to improve FC data visualization by capturing multidimensionality and integration of FC findings. Method: We analysed 12 normal/reactive bone marrow (N/R BM) samples and 12 BM samples from patients with myelodysplasia (MDS) with 10-color FC. All identifiable cell clusters were individually marked, grouped, and visualized on radar plots. RPs were optimized to de-clutter the cell clusters and map BM cell composition and maturation. Results: A total of 27 immature and mature cell clusters were identified and visualized on 8 multidimensional radar plots. The RPs displayed flow cytometry findings of normal BM in an integrated fashion to maximize overall insight into the data set. The constructed map of bone marrow cell composition was reproducible in all normal BM samples analyzed. Analysis of the pilot cohort of patient samples confirmed the presence of MDS-related changes. These changes are readily identifiable on RPs. Conclusion: We demonstrated that the cell clusters of normal BM can be mapped on multidimensional radar plots, which provide an inclusive insight into BM cell composition and maturation. These reproducible RPs present a comprehensive and comprehensible visual display of differentiation and maturation of haematopoietic cells in normal BM, and can be used as a reference map to assess abnormal haematopoiesis in MDS.
|
|
| 9. |
- Mahdi, Talal, et al.
(författare)
-
Characteristics of Lymphoproliferative Disorders with More Than One Aberrant Cell Population as Detected by 10-Color Flow Cytometry
- 2018
-
Ingår i: Cytometry Part B - Clinical Cytometry. - : Wiley. - 1552-4949. ; 94:2, s. 230-238
-
Tidskriftsartikel (refereegranskat)abstract
- Background: We have evaluated the frequency of lymphoproliferative disorders with more than one aberrant population of monotypic B-cells detected during routine hematopathological diagnostics. Materials and Methods: 2600 samples peripheral (blood, bone marrow, fine-needle aspirate, lymph node, and pleural fluid cell suspensions) were analyzed using a 10-color B-cell panel and a 10-color T-cell panel. A 10-color plasma cell/lymphoplasmacytic panel was performed when appropriate. Results: 790/2600 samples (30%) showed at least one aberrant B-cell population and 27(1%) showed an aberrant T-cell population. 41/790 samples (5.1%) showed two aberrant B-cell populations. Thirteen patients had two B-cell populations with different surface immunoglobulin restriction (one kappa+ and one lambda+), most with B-cell chronic lymphocytic leukemia-related phenotype. Five cases showed two B-cell populations with the same light chain restriction but distinctly different immunophenotypes. In 23 cases, two populations had the same light chain restriction and differed by expression of one or 2 markers, thus, a possibility of intraclonal differentiation could not be excluded. Cases with possible intraclonal differentiation had a significantly higher proportion of aberrant B-cells than those with two coexisting aberrant B-cell populations (49.9% vs. 27.7%, p = 0.008). In only one sample one population of clonal B-cell and one clonal T-cell population with large granular lymphocyte related phenotype were found. Conclusion: Using our panels 5.1% of cases with lymphoproliferative disorder-associated aberrant findings show two aberrant (clonal) lymphoid and/or plasma cell populations.
|
|
| 10. |
- Roschupkina, Teona, et al.
(författare)
-
Subpopulations of T regulatory cells in blood stem cell harvests influence development of acute graft versus host disease in allogeneic transplant recipients
- 2018
-
Ingår i: Cytometry Part B - Clinical Cytometry. - : Wiley. - 1552-4949. ; 94:2, s. 264-269
-
Tidskriftsartikel (refereegranskat)abstract
- Background: CD4+ FoxP3+ regulatory T cells (Tregs) are the potent suppressors of activation and proliferation of conventional T cells. Tregs subdivided by their expression of FoxP3 and CD45RA identify clinically important functional subsets. Methods: We analyzed Treg subpopulations in hematopoietic stem cell harvests (SCH) from 22 allogeneic (matched unrelated and sibling) donors with flow cytometry by their expression of CD45RA, CD127, CD25, and FoxP3 marker combinations. Results: A high fraction of "activated Tregs", defined as CD4+ FoxP3hiCD45RAlo (aTreg) cells relative to all CD4+ T-cells, in the SCH correlated with no subsequent development of acute graft-versus-host disease (aGVHD) in the corresponding transplant recipients (aTreg 1.29%, range 0.96-1.64%, vs. 0.23%, range 0.14-0.56%, with subsequent aGvHD; P = 0.0015). The "non-Treg" cells, defined by CD4+ FoxP3med/loCD45RAlo, and resting Treg (rTreg) cells, defined by CD4+ FoxP3medCD45RAhi, did not correlate with aGvHD development. We also showed that phenotypic aTregs could be induced in vitro from nonTregs under homeostatic proliferation conditions and that this ability correlated with the CD127 and CD25 expression patterns. Conclusions: We identified a subset of T CD4+ FoxP3+ cells, i.e., aTregs that were correlated to aGvHD development, and demonstrated plasticity of the nonTreg population to provide phenotypic aTregs. This could have both a predictive clinical relevance in inflammatory conditions as well as support a rationale for development of cell targeted therapy.
|
|
