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flow cytometry immunophenotyping

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https://www.readbyqxmd.com/read/29161617/early-pregnancy-immune-biomarkers-in-peripheral-blood-may-predict-preeclampsia
#1
M D Salazar Garcia, Y Mobley, J Henson, M Davies, A Skariah, S Dambaeva, A Gilman-Sachs, K Beaman, C Lampley, J Kwak-Kim
We performed a prospective cohort study in 197 pregnant women. Peripheral blood was collected between 5 and 16 weeks of gestation. Intracellular cytokine analysis and immunophenotype were performed by flow-cytometry. Serum levels of cytokines and chemokines were analyzed by multiplex assay. 86 patients were eligible for the analysis and 10.5% (n=9) developed preeclampsia. Patients with preeclampsia had significantly higher percentage of CD3(+)CD4(+)TNFα(+) T helper (Th) 1 cells (45.4±10.3 vs 37.1±8.5, P=0...
November 2, 2017: Journal of Reproductive Immunology
https://www.readbyqxmd.com/read/29155042/imaging-flow-cytometry-to-assess-chromosomal-abnormalities-in-chronic-lymphocytic-leukaemia
#2
Henry Hui, Kathryn A Fuller, Hun Chuah, James Liang, Hasib Siddiqi, Dejan Radeski, Wendy N Erber
Chronic Lymphocytic Leukaemia (CLL), the most common leukaemia in the Western world, has a characteristic phenotype and prognosis largely defined by the presence of cytogenetic aberrations. The gold standard for detecting these cytogenetic abnormalities is interphase fluorescence in situ hybridisation (FISH) performed on cell smears or tissue sections on glass slides. Fluorescently labelled DNA probes bind to specific chromosomal regions and the signal detected by fluorescent microscopy. Generally only 200 cells are assessed and the limit of sensitivity is 3% positive cells...
November 15, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/29155023/therapy-related-myeloid-neoplasm-in-an-18-year-old-boy-with-b-lymphoblastic-leukemia
#3
Xin Qing, Eduard Panosyan, Changjun Yue, Ping Ji, Moran Gotesman, Samuel French, Junchao Cai
BACKGROUND: Acute lymphoblastic leukemia (ALL) is the most common pediatric malignancy. Acute myeloid leukemia or myelodysplastic syndrome during the course of ALL is a rare entity. Some of these cases are therapy-related while the others occur due to lineage switch. The correct diagnosis relies on comparing the immunophenotypes and cytogenetic/molecular alterations of the myeloid neoplasm and the ALL. We present the clinical, pathologic and cytogenetic features of a case of an 18-year-old male with prior treatment for B-lymphoblastic leukemia (B-ALL) who developed therapy-related myeloid neoplasm (t-MN) 4-5years after his initial diagnosis of B-ALL...
November 16, 2017: Experimental and Molecular Pathology
https://www.readbyqxmd.com/read/29154914/how-to-make-usage-of-the-standardized-euroflow-8-color-protocols-possible-for-instruments-of-different-manufacturers
#4
Michaela Nováková, Hana Glier, Naděžda Brdičková, Marcela Vlková, Ana Helena Santos, Margarida Lima, Mikael Roussel, Juan Flores-Montero, Tomasz Szczepanski, Sebastian Böttcher, Vincent H J van der Velden, Paula Fernandez, Ester Mejstříková, Leire Burgos, Bruno Paiva, Jacques J M van Dongen, Alberto Orfao, Tomáš Kalina
A critical component of the EuroFlow standardization of leukemia/lymphoma immunophenotyping is instrument setup. Initially, the EuroFlow consortium developed a step-by-step standard operating protocol for instrument setup of ≥8-color flow cytometers that were available in 2006, when the EuroFlow activities started. Currently, there are 14 instruments from 9 manufacturers capable of 3-laser excitation and ≥8 color measurements. The specific adaptations required in the instrument set-up to enable them to acquire the standardized 8-color EuroFlow protocols are described here...
November 15, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29143491/fine-needle-aspiration-evaluation-of-pancreatic-lymphoma-a-retrospective-study-of-25-cases-in-a-single-institution
#5
Jiehao Zhou, Howard Wu, Jingmei Lin, Xiaoyan Wang, Shanxiang Zhang, Harvey Cramer, Shaoxiong Chen
BACKGROUND: Accurate diagnosis of pancreatic lymphoma is crucial for clinical management. We evaluate the role of fine-needle aspiration (FNA) in the diagnosis of pancreatic lymphoma with the aid of flow cytometry and/or immunohistochemistry on the cell block. METHODS: Cases of pancreatic lymphoma were collected by searching our pathology laboratory information system over a period of 16 years. The clinical findings, cytologic features, and immunophenotypic results were reviewed...
November 16, 2017: Diagnostic Cytopathology
https://www.readbyqxmd.com/read/29140408/philadelphia-chromosome-like-mixed-phenotype-acute-leukemia-demonstrating-p2ry8-crlf2-fusion-and-jak1-mutation
#6
Sarah M Choi, John K Frederiksen, Charles W Ross, Dale L Bixby, Lina Shao
Objectives: Philadelphia chromosome-like (Ph-like) genetic alterations define a subset of B lymphoblastic leukemia/lymphoma (B-ALL), which represents a separate provisional entity in the World Health Organization 2016 updated classification. However, these alterations have not been described outside the context of B-ALL. Methods: Cytogenomic array and molecular analysis identified a Ph-like signature in a mixed-phenotype acute leukemia (MPAL), B/myeloid, confirmed using conventional immunophenotypic and cytochemical analysis...
November 11, 2017: American Journal of Clinical Pathology
https://www.readbyqxmd.com/read/29128070/flow-cytometric-assessment-of-chronic-myeloid-neoplasms
#7
REVIEW
Min Shi, Phuong Nguyen, Dragan Jevremovic
Flow cytometry immunophenotyping of the hematopoietic cells from the bone marrow can help with diagnosis, prognosis, and therapy of chronic myeloid neoplasms. Unlike with B-cell neoplasms, there is no simple phenotypic test to substitute for clonality. Therefore, antigen panels to evaluate myeloid neoplasms are larger, and the gating strategies more complex than for lymphoid neoplasms. The number of phenotypic abnormalities in hematopoietic cells correlates with disease severity and cytogenetic complexity, and can be integrated into a scoring system for diagnostic and prognostic purposes...
December 2017: Clinics in Laboratory Medicine
https://www.readbyqxmd.com/read/29128068/b-lymphoblastic-leukemia-minimal-residual-disease-assessment-by-flow-cytometric-analysis
#8
REVIEW
Aaron C Shaver, Adam C Seegmiller
Among the most thoroughly evaluated modalities for assessment of minimal residual disease (MRD) in B acute lymphoblastic leukemia is multiparameter flow cytometry. Flow cytometric evaluation of MRD for B-ALL requires complete understanding of the immunophenotype of hematogones, the normal counterpart of leukemic B lymphoblasts. Assessment of multiple flow cytometry markers, in concert with each other in multidimensional histograms, is necessary to distinguish hematogones from malignant blasts. Emerging therapies targeting CD19 and other B-cell markers can disrupt the most frequently MRD assessment, requiring a revised approach as use of targeted therapies becomes widespread...
December 2017: Clinics in Laboratory Medicine
https://www.readbyqxmd.com/read/29128067/acute-myeloid-leukemia-immunophenotyping-by-flow-cytometric-analysis
#9
REVIEW
Xueyan Chen, Sindhu Cherian
Flow cytometry plays an indispensible role in the diagnosis and subclassification of acute myeloid leukemia (AML). Using a multiparametric approach, flow cytometry immunophenotyping has the advantage of efficiency with high sensitivity. This article reviews the general gating strategy, antibody panels for routine analysis, and additional markers for lineage assignment in the subclassification of AML. Also discussed are diagnostic immunophenotypic features of hard-to-classify entities considered within the differential diagnosis of AML...
December 2017: Clinics in Laboratory Medicine
https://www.readbyqxmd.com/read/29128066/flow-cytometry-of-t%C3%A2-cells-and-t-cell-neoplasms
#10
REVIEW
Jeffrey W Craig, David M Dorfman
Flow cytometry is ideally suited for the immunophenotypic analysis of T-cell neoplasia. This article covers the spectrum of flow cytometric findings associated with frequently encountered benign and neoplastic T-cell populations and details the most common immunophenotypic features associated with specific neoplasms of both immature and mature T cells.
December 2017: Clinics in Laboratory Medicine
https://www.readbyqxmd.com/read/29128065/flow-cytometry-of-b-cell-neoplasms
#11
REVIEW
Steven H Kroft, Alexandra M Harrington
Flow cytometric evaluation is considered a standard ancillary study for the diagnosis of most B-cell lymphoproliferative disorders. Establishing a neoplastic B-cell population depends on identification of light chain restriction or lack of light chain expression in mature neoplasms and demonstration of aberrant antigen expression in both immature and mature neoplasms, as compared with normal counterparts. The immunophenotypes of the most common B-cell neoplasms are herein described, with an emphasis on their immunophenotypic differential diagnosis and prognostic and therapeutic implications...
December 2017: Clinics in Laboratory Medicine
https://www.readbyqxmd.com/read/29119116/use-of-limiting-dilution-method-for-isolation-of-nucleus-pulposus-mesenchymal-stem-progenitor-cells-and-effects-of-plating-density-on-biological-characteristics-and-plasticity
#12
Linghan Lin, Zhiwei Jia, Yachao Zhao, Yaohong Wu, Xiyan Zhao, Yong Li, Ziming Guo, Jiahai Chen, Shi Cheng, Deli Wang, Dike Ruan
Objectives: To evaluate the effects of the limiting dilution method and plating density in rat nucleus pulposus mesenchymal stem/progenitor cells (NPMSCs). Materials and Methods: Nucleus pulposus tissues were isolated from 12-week-old male Sprague-Dawley rats and NPMSCs were isolated using limiting dilution method. Cells were then classified into 3 groups according to plating density. Cell morphologies were observed, and colony-forming units, migration abilities, proliferative capacities, cell cycle percentages, multilineage differentiation capacities, stem cell biomarker expression levels, and immunophenotyping were also examined in each group...
2017: BioMed Research International
https://www.readbyqxmd.com/read/29117436/isolation-cryopreservation-and-immunophenotyping-of-human-peripheral-blood-mononuclear-cells
#13
Fredine T Lauer, Jesse L Denson, Scott W Burchiel
This unit describes procedures for the isolation, cryopreservation, and thawing of human peripheral blood mononuclear cells (HPBMC) and analysis of cell surface markers (CSM) for immunophenotyping using polychromatic flow cytometry. This methodology can be used to ensure that cell integrity and phenotype stability are not altered through cryopreservation and extended storage. For this analysis, HPBMC were isolated from 7 healthy individuals, and 11-color flow cytometry was performed on freshly isolated samples as well as samples cryopreserved for short- and long-term periods...
November 8, 2017: Current Protocols in Toxicology
https://www.readbyqxmd.com/read/29115725/comparison-of-conventional-cytomorphology-flow-cytometry-immunophenotyping-and-automated-cell-counting-of-csf-for-detection-of-cns-involvement-in-acute-lymphoblastic-leukemia
#14
J C Jaime-Pérez, M F Borrego-López, R A Jiménez-Castillo, N Méndez-Ramírez, R Salazar-Riojas, L T Fernández, D Gómez-Almaguer
BACKGROUND AND OBJECTIVE: Cytospin conventional cytomorphology (CCC) is the standard method for detecting lymphoblasts in cerebrospinal fluid (CSF) of patients with acute lymphoblastic leukemia (ALL) and for guiding treatment decisions. We evaluated flow cytometry immunophenotyping (FCI) performance for improving detection of central nervous system (CNS) involvement in ALL. METHODS: This prospective study included analysis of consecutive CSF samples of patients of all ages with ALL at 3 clinical stages: new diagnosis, relapse suspicion, and after relapse treatment...
November 8, 2017: International Journal of Laboratory Hematology
https://www.readbyqxmd.com/read/29113330/flow-cytometric-analysis-of-cd64-expression-pattern-and-density-in-the-diagnosis-of-acute-promyelocytic-leukemia-a-multi-center-study-in-shanghai-china
#15
Min Liu, Xiangqin Weng, Shenglan Gong, Hui Chen, Jing Ding, Mengqiao Guo, Xiaoxia Hu, Jianmin Wang, Jianmin Yang, Gusheng Tang
No unified immunophenotypic profiles and corresponding analytic strategies have been established for the rapid diagnosis of acute promyelocytic leukemia (APL) using flow cytometry (FCM). Here we describe a characteristic immunophenotypic panel that can rapidly and accurately distinguish APL from other types of adult acute myeloid leukemia (AML) using only FCM. By comparing APL cells and non-APL AML cells that share APL common immunophenotypes (CD34(-)CD117(+)HLA(-)DR(-)) we found that CD64 was a significant factor that differentiated APL from other AMLs...
October 6, 2017: Oncotarget
https://www.readbyqxmd.com/read/29103430/promise-progress-and-pitfalls-in-the-search-for-central-nervous-system-biomarkers-in-neuroimmunological-diseases-a-role-for-cerebrospinal-fluid-immunophenotyping
#16
Bibiana Bielekova, Michael R Pranzatelli
Biomarkers are central to the translational medicine strategic focus, though strict criteria need to be applied to their designation and utility. They are one of the most promising areas of medical research, but the "biomarker life-cycle" must be understood to avoid false-positive and false-negative results. Molecular biomarkers will revolutionize the treatment of neurological diseases, but the rate of progress depends on a bold, visionary stance by neurologists, as well as scientists, biotech and pharmaceutical industries, funding agencies, and regulators...
August 2017: Seminars in Pediatric Neurology
https://www.readbyqxmd.com/read/29101404/autoreactive-t-cells-in-immunopathogenesis-of-tb-associated-uveitis
#17
Ravichandra Tagirasa, Shaifaly Parmar, Manas R Barik, Satish Devadas, Soumyava Basu
Purpose: Intraocular inflammation in tuberculosis-associated uveitis (TBU) is usually widespread, and responds unpredictably to treatment. Herein, we analyze the intraocular T-cell response in TBU for its surface phenotype, antigenic specificity, and functional characteristics to explain the above observations. Methods: We isolated T cells from vitreous humor samples of patients with TBU and non-TB uveitis (controls). These were directly stained for surface markers CD4, CD8, CD45RO, CD45RA, CCR7, as well as intracellular cytokines IFN-γ, TNF-α, and IL-17 and analyzed on flow cytometry...
November 1, 2017: Investigative Ophthalmology & Visual Science
https://www.readbyqxmd.com/read/29094769/differentiation-and-characterization-of-rat-adipose-tissue-mesenchymal-stem-cells-into-endothelial-like-cells
#18
V Cannella, G Piccione, R Altomare, A Marino, P Di Marco, L Russotto, S Di Bella, G Purpari, F Gucciardi, G Cassata, G Damiano, V D Palumbo, A Santoro, C Russo Lacerna, A I Lo Monte, A Guercio
In this study, mesenchymal stem cells were isolated from rat adipose tissue (AD-MSCs) to characterize and differentiate them into endothelial-like cells. AD-MSCs were isolated by mechanical and enzymatic treatments, and their identity was verified by colony-forming units (CFU) test and by differentiation into cells of mesodermal lineages. The endothelial differentiation was induced by plating another aliquot of cells in EGM-2 medium, enriched with specific endothelial growth factors. Five subcultures were performed...
November 2, 2017: Anatomia, Histologia, Embryologia
https://www.readbyqxmd.com/read/29071684/immunophenotypic-identification-of-early-myeloerythroid-development
#19
Cornelis J H Pronk, David Bryder
Myeloerythroid-restricted precursor cells, derived from multipotent hematopoietic stem cells, give rise to mature cells of the granulocyte, monocyte, erythroid, and/or thrombocytic lineages. High-resolution profiling of the developmental stages, from hematopoietic stem cells to mature progeny, is important to be able to study and understand the underlying mechanisms that guide various cell fate decisions. Also, this approach opens for greater insights into pathogenic events such as leukemia, diseases that are most often characterized by halted differentiation at defined immature precursor levels...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29071677/quantitative-fluorescence-measurements-with-multicolor-flow-cytometry
#20
Lili Wang, Adolfas K Gaigalas, James Wood
Multicolor flow cytometer assays are routinely used in clinical laboratories for immunophenotyping, monitoring disease and treatment, and determining prognostic factors. However, existing methods for quantitative measurements have not yet produced satisfactory results independent of flow cytometers used. This chapter details a procedure for quantifying surface and intracellular protein biomarkers by calibrating the output of a multicolor flow cytometer in units of antibodies bound per cell (ABC). The procedure includes the following critical steps: (a) quality control (QC) and performance characterization of the multicolor flow cytometer, (b) fluorescence calibration using hard dyed microspheres assigned with fluorescence intensity values in equivalent number of reference fluorophores (ERF), (c) compensation for correction of fluorescence spillover, and (d) application of a biological reference standard for translating the ERF scale to the ABC scale...
2018: Methods in Molecular Biology
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