Flow cytometry immunophenotypic analysis of Philadelphia-negative myeloproliferative neoplasms: Correlation with histopathologic features

2014 ◽  
pp. n/a-n/a ◽  
Author(s):  
Juan Ouyang ◽  
Wenli Zheng ◽  
Qi Shen ◽  
Maitrayee Goswami ◽  
Jeffrey L. Jorgensen ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Myeloproliferative Neoplasms ◽  
Histopathologic Features ◽  
Immunophenotypic Analysis

U.S.-Canadian consensus recommendations on the immunophenotypic analysis of hematologic neoplasia by flow cytometry

Cytometry ◽  
1997 ◽  
Vol 30 (5) ◽  
pp. 213-213 ◽  
Author(s):  
Raul C. Braylan ◽  
Michael J. Borowitz ◽  
Bruce H. Davis ◽  
Gregory T. Stelzer ◽  
Carleton C. Stewart
Keyword(s):  
Flow Cytometry ◽  
Consensus Recommendations ◽  
Hematologic Neoplasia ◽  
Immunophenotypic Analysis

scholarly journals Immunophenotypic Analysis of Anaplastic Large Cell Lymphoma by Flow Cytometry

2003 ◽  
Vol 119 (2) ◽  
pp. 205-212 ◽  
Author(s):  
Jonathan Juco ◽  
Jeannine T. Holden ◽  
Karen P. Mann ◽  
Lloyd G. Kelley ◽  
Shiyong Li
Keyword(s):  
Flow Cytometry ◽  
Anaplastic Large Cell Lymphoma ◽  
Cell Lymphoma ◽  
Large Cell ◽  
Large Cell Lymphoma ◽  
Immunophenotypic Analysis

scholarly journals Multiparameter flow cytometry reveals myelodysplasia-related aberrant antigen expression in myelodysplastic/myeloproliferative neoplasms

2013 ◽  
Vol 84B (3) ◽  
pp. 194-197 ◽  
Author(s):  
Wolfgang Kern ◽  
Ulrike Bacher ◽  
Susanne Schnittger ◽  
Tamara Alpermann ◽  
Claudia Haferlach ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Myeloproliferative Neoplasms ◽  
Antigen Expression ◽  
Multiparameter Flow Cytometry

scholarly journals The potential of proliferative and apoptotic parameters in clinical flow cytometry of myeloid malignancies

2021 ◽  
Vol 5 (7) ◽  
pp. 2040-2052
Author(s):  
Stefan G. C. Mestrum ◽  
Anton H. N. Hopman ◽  
Frans C. S. Ramaekers ◽  
Math P. G. Leers
Keyword(s):  
Flow Cytometry ◽  
Myeloproliferative Neoplasms ◽  
Diagnostic Process ◽  
Multiparameter Flow Cytometry ◽  
Major Step ◽  
Differentiation Markers ◽  
Myeloid Malignancies ◽  
Cell Lineages ◽  
Apoptotic Markers ◽  
Multiparameter Analysis

Abstract Standardization of the detection and quantification of leukocyte differentiation markers by the EuroFlow Consortium has led to a major step forward in the integration of flow cytometry into classification of leukemia and lymphoma. In our opinion, this now enables introduction of markers for more dynamic parameters, such as proliferative and (anti)apoptotic markers, which have proven their value in the field of histopathology in the diagnostic process of solid tumors and lymphoma. Although use of proliferative and (anti)apoptotic markers as objective parameters in the diagnostic process of myeloid malignancies was studied in the past decades, this did not result in the incorporation of these biomarkers into clinical diagnosis. This review addresses the potential of these markers for implementation in the current, state-of-the-art multiparameter analysis of myeloid malignancies. The reviewed studies clearly recognize the importance of proliferation and apoptotic mechanisms in the pathogenesis of bone marrow (BM) malignancies. The literature is, however, contradictory on the role of these processes in myelodysplastic syndrome (MDS), MDS/myeloproliferative neoplasms, and acute myeloid leukemia. Furthermore, several studies underline the need for the analysis of the proliferative and apoptotic rates in subsets of hematopoietic BM cell lineages and argue that these results can have diagnostic and prognostic value in patients with myeloid malignancies. Recent developments in multiparameter flow cytometry now allow quantification of proliferative and (anti)apoptotic indicators in myeloid cells during their different maturation stages of separate hematopoietic cell lineages. This will lead to a better understanding of the biology and pathogenesis of these malignancies.

Derangements of Toll-like Receptors, Inflammatory Cytokines, and Reactive Oxygen Species in Philadelphia Chromosome-Negative Myeloproliferative Neoplasm: Implicate Roles of Inflammation in the Pathogenesis

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 1945-1945 ◽  
Author(s):  
Guanfang Shi ◽  
Hui Chen ◽  
Cherif Abdelmalek ◽  
Andrei Bandarchuk ◽  
Abdullah Khawer Mahmood ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Dendritic Cells ◽  
Inflammatory Cytokines ◽  
Mononuclear Cells ◽  
Conflicts Of Interest ◽  
Myeloproliferative Neoplasms ◽  
Philadelphia Chromosome ◽  
Toll Like Receptors ◽  
Clonal Proliferation ◽  
Inflammatory Processes

Abstract Philadelphia chromosome-negative myeloproliferative neoplasms (MPN) including essential thrombocythemia(ET), polycythemia vera(PV), and myelofibrosis (MF) have been regarded as a stem cell diseases with malignant clonal proliferation; but recently, inflammatory processes have been proposed as playing found a major role in the pathogenesis of MPN. Toll-like receptors (TLRs) are a family of pattern-recognition receptors that function as key initiators of innate immunity signaling, then induce inflammatorycytokines, and ROS formation. Therefore we measured TLRs, inflammatory cytokines, and ROS in patients with MPN to assess the role of inflammation in MPN. Methods: TLR assay.TLR-2, 4, 7, 9 quantification was performed by immuno-staining of 1×106 mononuclear cells which were incubated with fluorescence-conjugated anti-TLR-2, 4, 7, 9 antibodies and assayed by flow cytometry. Monocytes culture and dendritic cells differentiation.Human monocytes were isolated from human peripheral blood mononuclear cells (PBMNC) using Pan Monocyte Isolation Kit. Isolated monocytes were incubated with IL-4 and GM-CSF in cultures to differentiate to dendric cells . Immature dendritic cells were either left untreated or stimulated with Pam3CSK4. The maturation of dendritic cells was determined by flow cytometry using CD80, CD83, CD11c, and HLA-DR. Multiplex ELISA. Human plasma and cell culture supernatants were analyzed in duplicates by Meso Scale Discovery Multi-Spot Assay system. In total, ten cytokines were assayed: IFN-g, IL-1b, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13, and TNF-a. Cell ROS measurement. Cellular ROS was determined by a dichlorofluorescein (DCF) assay. PBMNCs were incubated with 10 µM 2′,7′-dichlorodihydrofluorescein diacetate (carboxy-H2DCFDA) (Life Technology) for 30 min at 37°C. The oxidation of H2DCFDA was measured and analyzed by flowcytometery. Results. 1)TLR2 was the only TLR found to be elevated in PV or ET patients (Fig 1). 2) PlasmaIL-1b levels were elevated in TLR2 high patients than TLR-2 low patients. 3) No difference between TLR-2 high and low patients in the maturation of monocytes to dendritic cells. 4) Monocyte derived dendritic cells with high TLR-2 patientsreleased more IL-8 and TNF-a after Pam3CSK4 stimulation (Fig 2). 5) ROS were more elevated in MF patients than PV, ET patients, and controls (Fig 3). Conclusion. 1) TLR 2 is significantly elevated in PV and some ET patients and TLR-2 high patients were found to have elevated plasma IL-1b,and IL-8, and TNF-α in monocytes-derived dendric cell cultures afterPam3CSK4 stimulation than TLR-2 low patients. This confirms that TLR-2 is deranged in PV and ET. 2) ROS is elevated in MF patients compared to ET and PV patients, or controls. Thus, this study suggests that inflammatory processes likely play a role in the pathogenesis of Ph- MPN through first TLR-2 derangement in PV and ET , then through years of chronic inflammatory process , with the accumulation of more ROS seen in MF, which caused more damage to the DNA resulting more malignant clonal proliferation. A similar phenomenon was observed that in JAK2V617F mutation , allele-burden was observed gradually increased from ET , PV then to MF . Disclosures No relevant conflicts of interest to declare.

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