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Immunophenotyping by flow cytometry

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https://www.readbyqxmd.com/read/29143491/fine-needle-aspiration-evaluation-of-pancreatic-lymphoma-a-retrospective-study-of-25-cases-in-a-single-institution
#1
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/29128068/b-lymphoblastic-leukemia-minimal-residual-disease-assessment-by-flow-cytometric-analysis
#2
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
#3
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/29117436/isolation-cryopreservation-and-immunophenotyping-of-human-peripheral-blood-mononuclear-cells
#4
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
#5
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
#6
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
#7
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
#8
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
#9
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
#10
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
#11
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
https://www.readbyqxmd.com/read/29058101/dock8-deficiency-presenting-as-an-ipex-like-disorder
#12
Fayhan J Alroqi, Louis-Marie Charbonnier, Sevgi Keles, Fatima Ghandour, Pierre Mouawad, Rami Sabouneh, Reem Mohammed, Abduarahman Almutairi, Janet Chou, Michel J Massaad, Raif S Geha, Zeina Baz, Talal A Chatila
PURPOSE: The dedicator of cytokinesis 8 (DOCK8) deficiency is an autosomal recessive-combined immunodeficiency whose clinical spectra include recurrent infections, autoimmunity, malignancies, elevated serum IgE, eczema, and food allergies. Here, we report on patients with loss of function DOCK8 mutations with profound immune dysregulation suggestive of an immune dysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX)-like disorder. METHODS: Immunophenotyping of lymphocyte subpopulations and analysis of DOCK8 protein expression were evaluated by flow cytometry...
November 2017: Journal of Clinical Immunology
https://www.readbyqxmd.com/read/29057538/enhanced-gastric-cancer-growth-potential-of-mesenchymal-stem-cells-derived-from-gastric-cancer-tissues-educated-by-cd4-t-cells
#13
Rongman Xu, Xiangdong Zhao, Yuanyuan Zhao, Bin Chen, Li Sun, Changgen Xu, Bo Shen, Mei Wang, Wenrong Xu, Wei Zhu
OBJECTIVES: Gastric cancer mesenchymal stem cells (GC-MSCs) can promote the development of tumour growth. The tumour-promoting role of tumour-associated MSCs and T cells has been demonstrated. T cells as the major immune cells may influence and induce a pro-tumour phenotype in MSCs. This study focused on whether CD4(+) T cells can affect GC-MSCs to promote gastric cancer growth. MATERIALS AND METHODS: CD4(+) T cells upregulation of programmed death ligand 1 (PD-L1) expression in GC-MSCs through the phosphorylated signal transducer and activator of transcription (p-STAT3) signalling pathway was confirmed by immunofluorescence, western blotting and RT-PCR...
October 22, 2017: Cell Proliferation
https://www.readbyqxmd.com/read/29045966/-immune-effects-of-specific-ctls-response-induced-by-dendritic-cells-pulsed-with-ny-eso-1-peptide
#14
J W Liu, X Lu, Z M Yang, L J Deng, L Yang
OBJECTIVE: To explore the potential of autologous dendritic cells (DCs) pulsed with caner/testis antigen NY-ESO-1 peptides in inducing specific cytotoxic T lymphocyte (CTLs) response and antineoplastic immune function of specific CTLs. METHODS: Fifteen patients with II to III stage positive HLA -A0201(+) and NY-ESO-1(+) were enrolled in the Cancer Hospital Chinese Academy of Medical Sciences on the basis of preclinical experiments from November 2014 to October 2015, and their peripheral blood mononuclear cells (PBMCs) and peripheral blood lymphocytes (PBLs) were isolated...
October 18, 2017: Beijing da Xue Xue Bao. Yi Xue Ban, Journal of Peking University. Health Sciences
https://www.readbyqxmd.com/read/29035453/immunophenotyping-of-chronic-lymphocytic-leukemia
#15
Mesude Falay And Gulsum Ozet
BACKGROUND: Chronic Lymphocytic Leukemia (CLL) is one of the most common diagnoses made by flow cytometry laboratories. There is no consensus on which markers need to be used in flow cytometry for accurate immunophenotyping. Herein, we investigated the role of markers used in flow cytometry in the distinction between CLL and MCL. METHODS: A total 339 recently diagnosed B lymphoproliferative patient cases were retrospectively studied for their immunophenotypical propoerties using flow cytometry...
October 1, 2017: Clinical Laboratory
https://www.readbyqxmd.com/read/29031042/humoral-and-cellular-immune-response-of-mice-challenged-with-yersinia-pestis-antigenic-preparations
#16
Elida A Leal, Josimar D Moreira, Fernanda F Nunes, Larissa R Souza, Janaina M Martins, Vicente P C Toledo, Alzira M P Almeida, Tania M P Guimarães
OBJECTIVES: The plague, which is an infectious disease caused by Yersinia pestis, still threatens many populations in several countries. The worldwide increase in human plague cases and the potential use of the bacteria as a biological weapon reinforce the need to study the immunity that is induced by potential vaccine candidates. To determine the immunogenicity of antigenic preparations based on the F1 protein and the total extract from Y. pestis, we assessed the role of these antigens in inducing an immune response...
October 12, 2017: Brazilian Journal of Infectious Diseases
https://www.readbyqxmd.com/read/29030811/flow-cytometric-detection-of-g0-in-live-cells-by-hoechst-33342-and-pyronin-y-staining
#17
Ayad Eddaoudi, Stephanie Louise Canning, Itaru Kato
Hoechst 33342 and Pyronin Y double staining can be used to measure DNA and RNA content in live cells by flow cytometry. Quiescent cells at G0 phase have the same amount of DNA as cells at G1 phase but lower RNA levels compared to proliferating cells. Therefore, resting cells in G0 phase can be distinguished from proliferating cells in G1, S, and G2 M phases. This chapter describes a protocol for double staining of live cells with Hoechst 33342 and Pyronin Y. Combined with immunophenotyping of intact and live cells Hoechst 33342 and Pyronin Y staining is a powerful noninvasive method for the analysis and isolation of quiescent cells from any defined cell population...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29027383/immunological-prediction-of-cytomegalovirus-cmv-replication-risk-in-solid-organ-transplantation-recipients-approaches-for-regulating-the-targeted-anti-cmv-prevention-strategies
#18
REVIEW
Sang Hoon Han
The current cytomegalovirus (CMV) prevention strategies in solid organ transplantation (SOT) recipients have contributed towards overcoming the detrimental effects caused by CMV lytic infection, and improving the long-term success rate of graft survival. Although the quantification of CMV in peripheral blood is the standard method, and an excellent end-point for diagnosing CMV replication and modulating the anti-CMV prevention strategies in SOT recipients, a novel biomarker mimicking the CMV control mechanism is required...
September 2017: Infection & Chemotherapy
https://www.readbyqxmd.com/read/29024461/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
#19
Andy C Rawstron, Karl-Anton Kreuzer, Asha Soosapilla, Martin Spacek, Olga Stehlikova, Peter Gambell, Neil McIver-Brown, Neus Villamor, Katherina Psarra, Maria Arroz, Raffaella Milani, Javier de la Serna, M Teresa Cedena, Ozren Jaksic, Josep Nomdedeu, Carol Moreno, Gian Matteo Rigolin, Antonio Cuneo, Preben Johansen, Hans E Johnsen, Richard Rosenquist, Carsten Utoft Niemann, Wolfgang Kern, David Westerman, Marek Trneny, Stephen Mulligan, Michael Doubek, Sarka Pospisilova, Peter Hillmen, David Oscier, Michael Hallek, Paolo Ghia, Emili Montserrat
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...
October 10, 2017: Cytometry. Part B, Clinical Cytometry
https://www.readbyqxmd.com/read/28991896/t-cell-regulation-in-erythema-nodosum-leprosum
#20
Edessa Negera, Stephen L Walker, Kidist Bobosha, Rawleigh Howe, Abraham Aseffa, Hazel M Dockrell, Diana N Lockwood
Leprosy is a disease caused by Mycobacterium leprae where the clinical spectrum correlates with the patient immune response. Erythema Nodosum Leprosum (ENL) is an immune-mediated inflammatory complication, which causes significant morbidity in affected leprosy patients. The underlying cause of ENL is not conclusively known. However, immune-complexes and cell-mediated immunity have been suggested in the pathogenesis of ENL. The aim of this study was to investigate the regulatory T-cells in patients with ENL...
October 2017: PLoS Neglected Tropical Diseases
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