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Cytometry. Part B, Clinical Cytometry

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We are pleased to present abstracts from ESCCA 2018: At the Shore of Future Cytometry. This year, the European Society for Clinical Cell Analysis (ESCCA) organised its annual Conference over 3 days in Valencia, Spain. Four workshops for a limited number of participants - dedicated to Cytometry Analysis Tools, Cytometry of Microparticles, Cytometry in Veterinary and Animal Sciences and CLL MRD - as well as ESCCA certification exams took place before the Conference. The Conference itself included four Education Programmes from which the participants had to choose in advance: (i) Good Cytometry Practices; (ii) Advanced Hemato-Oncology; (iii) Clinical Diagnostic Cytometry and (v) Advanced Immunology...
December 5, 2018: Cytometry. Part B, Clinical Cytometry
Fan Wu, Ying Pan, Huiping Wang, Qianshan Tao, Furun An, Jiakui Zhang, Zhimin Zhai
Hepatosplenic T-cell lymphoma (HSTCL) is a very rare non-Hodgkin lymphoma with an aggressive clinical course and poor prognosis. Patients of this disease usually presented with hepatosplenomegaly, which can be misdiagnosed or delayed. Bone marrow (BM) and peripheral blood (PB) are frequently involved, however, central nervous system (CNS) involvement is less common. Here, we are reporting an unusual case of hepatosplenic γδ T-cell lymphoma in a 64-year-old man with CNS involvement. Flow cytometry immunophenotyping was proved of great diagnostic contribution...
November 30, 2018: Cytometry. Part B, Clinical Cytometry
Titus Keller, Leonie Wengenroth, Denise Smorra, Kristina Probst, Leo Kurian, Angela Kribs, Bent Brachvogel
BACKGROUND: Human breast milk could be an important stem cell source for the development of newborn and preterm infants, but quantitative data on the stem cell content in breast milk at various gestational stages are needed to determine the clinical value of breast milk as a source of stem cells. Breast milk also contains milk fat globules, lipid droplets of different sizes, debris and dead cells and these components hamper flow cytometry analysis of human breast milk samples. METHODS: Here, we originally used standard protocols for flow cytometry to characterize cell populations in human breast milk but failed to discriminate between cells and noncellular components...
November 26, 2018: Cytometry. Part B, Clinical Cytometry
Claire Carrion, Estelle Guérin, Nathalie Gachard, Alexandre le Guyader, Stéphane Giraut, Jean Feuillard
BACKGROUND: The different B-cell subsets in human bone marrow result from a dynamic equilibrium between endogenous production, B-cell bone marrow reentry and terminal plasma cell differentiation. Our aim was to define and quantify the different medullary B-cell subsets. METHODS: A series of 32 normal adult bone marrows plus 15 normal adult blood samples was studied by nine color flow cytometry (CD10, CD19, CD24, CD27, CD34, CD38, CD45, IgM, and IgD). With the Kaluza software radar plots, two 2D triple parametric histograms (CD10/CD34/CD45 and CD27/IgM/IgD) were set-up to identify six progenitor and five mature B-cell subsets...
November 18, 2018: Cytometry. Part B, Clinical Cytometry
Anne E Bras, Valerie de Haas, Arthur van Stigt, Mojca Jongen-Lavrencic, H Berna Beverloo, Jeroen G Te Marvelde, C Michel Zwaan, Jacques J M van Dongen, Jeanette H W Leusen, Vincent H J van der Velden
BACKGROUND: While it is known that CD123 is normally strongly expressed on plasmacytoid dendritic cells and completely absent on nucleated red blood cells, detailed information regarding CD123 expression in acute leukemia is scarce and, if available, hard to compare due to different methodologies. METHODS: CD123 expression was evaluated using standardized EuroFlow immunophenotyping in 139 pediatric AML, 316 adult AML, 193 pediatric BCP-ALL, 69 adult BCP-ALL, 101 pediatric T-ALL, and 28 adult T-ALL patients...
November 18, 2018: Cytometry. Part B, Clinical Cytometry
Deepashri Rao, Manjunatha M Venkataswamy, Ravi Vasanthapuram, P Satishchandra, Anita Desai
BACKGROUND: Neurotuberculosis is one of the commonest HIV associated opportunistic infections of the central nervous system in India. HIV-TB coinfection may lead to altered frequencies of T cells, thereby influencing the course and progression of the disease. METHODS: We examined the frequencies of T cell subsets in HIV infected individuals with neurotuberculosis (HIV+nTB+) as compared to individuals with HIV associated systemic TB (HIV+sTB+), asymptomatic HIV (HIV+TB-), non-HIV neuro TB (HIV-nTB+), non-HIV systemic TB (HIV-sTB+), and healthy controls (HIV-TB-)...
November 18, 2018: Cytometry. Part B, Clinical Cytometry
Shovik Bandyopadhyay, Jared S Fowles, Liyang Yu, Daniel A C Fisher, Stephen T Oh
BACKGROUND: Background: Mass cytometry (CyTOF) is a powerful tool for analyzing cellular networks at the single cell level. Due to the high-dimensional nature of this approach, analysis algorithms have been developed to visualize and interpret mass cytometry data. In this study, we applied these approaches to a cohort of patients with secondary acute myeloid leukemia (sAML). METHODS: We utilized mass cytometry to interrogate localization and intensity of thrombopoietin-mediated intracellular signaling in sAML...
November 13, 2018: Cytometry. Part B, Clinical Cytometry
Guilhem Requirand, Nicolas Robert, Stéphanie Boireau, Laure Vincent, Anja Seckinger, Salaheddine Bouhya, Patrice Ceballos, Guillaume Cartron, Dirk Hose, Bernard Klein, Jérôme Moreaux
BACKGROUND: Mutiple myeloma (MM) is a neoplasia characterized by the accumulation of malignant plasma cells (PC) in the bone marrow. Although proliferation markers have been studied in MM, none of the current staging systems include them. Moreover, approaches used to analyze proliferation do not separate MM cells (MMCs) from normal PC. METHODS: In this study, we combined multiparameter flow cytometry and BrdU incorporation or Ki67 staining to analyze MM cell proliferation in 44 monoclonal gammopathy of undetermined significance (MGUS), 153 newly diagnosed MM patients and 69 MM patients at relapse...
November 12, 2018: Cytometry. Part B, Clinical Cytometry
Yi Zhou, Andres Moon, Eric Hoyle, Jonathan R Fromm, Xueyan Chen, Lori Soma, Stephen J Salipante, Brent L Wood, David Wu
BACKGROUND: The presence of measurable residual disease after therapy is a significant risk factor of relapse in patients with acute myeloid leukemia (AML). By detecting cells with leukemia-associated immunophenotype (LAIP), multiparameter flow cytometry (MFC) can detect residual leukemia at a level significantly lower than that detected by morphology. However, changes in LAIPs during or after therapy may pose a challenge to MRD testing. AML with mutated NPM1 represents the largest subtype of AML sharing a common leukemogenic mechanism and similar LAIPs...
November 12, 2018: Cytometry. Part B, Clinical Cytometry
Hector Sarmiento Palao, Fabián Tarín, Fernanda Martirena, Eva Barragán, Esperanza Such, Amparo Sempere, Maria Tasso, Pablo Manresa, Francisco López
No abstract text is available yet for this article.
October 24, 2018: Cytometry. Part B, Clinical Cytometry
Daniel Harrison, Rosalie Ward, Sarah Bastow, Andrew Parr, Susan Macro, Paul K Wallace
BACKGROUND: An interlaboratory study was performed to compare the performance of the 3 mL TransFix®/EDTA Vacuum Blood Collection Tube (TVT), with the 5 mL Cyto-Chex® BCT tube (BCT). Both devices are intended for collection and storage of whole blood specimens for immunophenotyping of leukocytes by flow cytometry for up to 14 days. METHODS: One site in the United States and two in the United Kingdom tested samples from 10 HIV positive patients and four healthy subjects for a total of 42 samples...
October 24, 2018: Cytometry. Part B, Clinical Cytometry
D Robert Sutherland, Rumina Musani
No abstract text is available yet for this article.
October 24, 2018: Cytometry. Part B, Clinical Cytometry
Péter Szerémy, Katalin Tauberné Jakab, Sándor Baráth, András Apjok, Kata Filkor, Zsolt Holló, János Márki-Zay, János Kappelmayer, Sándor Sipka, Péter Krajcsi, Gergely Toldi
BACKGROUND: MDR transporters are important biomarkers of drug resistance in cancer and in autoimmune conditions. We determined the MDR1, MRP1 and BCRP activity in CD3+ lymphocytes using a flow cytometry based method from 120 healthy volunteers in order to describe normal reference values of the activity of these transporters. The effects of gender and age were also determined. METHODS: The Solvo MDQ Kit™ was used for measurements. In this assay, fluorescent reporter substrates (Calcein-AM for MDR1 and MRP1 and mitoxantrone for BCRP, respectively) are trapped in the cytoplasm and pumped out by MDR proteins depending on the presence or absence of specific inhibitors (verapamil for MDR1 and MRP1, indomethacin for MRP1 and KO134 for BCRP, respectively), allowing for quantitative, standardized assessment...
October 23, 2018: Cytometry. Part B, Clinical Cytometry
M Wheeler, G White, S Brockie, M Dickson, R Weinkove
BACKGROUND: Bone marrow (BM) aspirate analysis by flow cytometry (FC) is a key hematopathologic technique but dry taps hinder diagnosis. We describe the utility and limitations of a method of mechanically disaggregating BM trephine biopsies for FC. METHOD: Trephine biopsies mechanically disaggregated for FC between 2010 and 2016 were reviewed. We recorded cell yield, pathological findings, and turnaround time. Where available, results of trephine FC were compared with aspirate FC...
October 22, 2018: Cytometry. Part B, Clinical Cytometry
Ravnit-Kaur Grewal, Manogari Chetty, Emmanuel-Akinola Abayomi, Ciprian Tomuleasa, Jonathan R Fromm
Hodgkin's lymphoma (HL) has a unique immunophenotype derived from immunohistochemistry (positive for CD15, CD30, and Pax-5; negative for CD3, CD20 in most cases, and CD45). The knowledge gained over recent years enables better diagnosis, prognosis, and treatment of HL. Flow cytometry as a tool for the diagnosis of classic HL has not been useful in the past due to the difficulty in isolating Reed-Sternberg cells as they are admixed in a rich inflammatory background which consists mainly of T cells, B cells, eosinophils, histiocytes, and plasma cells...
October 22, 2018: Cytometry. Part B, Clinical Cytometry
Marina Garcia-Prat, Daniel Álvarez-Sierra, Aina Aguiló-Cucurull, Sandra Salgado-Perandrés, Sara Briongos-Sebastian, Clara Franco-Jarava, Andrea Martin-Nalda, Roger Colobran, Isabel Montserrat, Manuel Hernández-González, Ricardo Pujol-Borrell, Pere Soler-Palacin, Mónica Martínez-Gallo
BACKGROUND: For the accurate diagnosis of immunodeficiencies is crucial to compare patients' immunology laboratory values with age-sex matched controls, yet there is a paucity of normal values for most populations. OBJECTIVES: To define appropriate reference values of extended lymphocyte subpopulations and T-cell receptor excision circle (TRECs) levels in healthy pediatric donors between 1 month and 18 years of age. METHODS: Extended immunophenotyping values were obtained by analysis of multiparameter flow cytometry panels for the following subpopulations: CD4+ and CD8+ Naive, Effector, Effector Memory and Central Memory, T helper subpopulations and their degrees of activation, T Regulatory cells, Recent Thymic Emigrants (RTE), B Lymphocyte subpopulations (Transitional, Naive, Preswitch-Memory, Switch-Memory, Plasmablasts, CD21low, and Exhausted), and subpopulations for Monocytes, NK cells and Dendritic Cells...
October 17, 2018: Cytometry. Part B, Clinical Cytometry
Ru Feng, Vijaya Raj Bhatt, Kai Fu, Samuel Pirruccello, Ji Yuan
OBJECTIVES: The purpose of this study was to determine whether immunophenotypic profiles detected by flow cytometry are useful in differentiating chronic myelomonocytic leukemia (CMML) from reactive monocytosis, and between CMML subtypes. METHODS: Eight-color flow cytometry was used to immunophenotype blasts, monocytes, and granulocytes in the bone marrow of 34 patients with CMML and 12 patients with reactive monocytosis. RESULTS: Bone marrow myeloblast, promonocyte, and monocyte counts by flow cytometry were significantly higher in the CMML group than in the reactive monocytosis group...
October 17, 2018: Cytometry. Part B, Clinical Cytometry
Nikolaos Gardikas, Myrofora Vikentiou, Evgenia Konsta, Christos K Kontos, Sotirios G Papageorgiou, Aris Spathis, Efthimia Bazani, Anthi Bouchla, Violetta Kapsimali, Katherina Psarra, Periklis Foukas, George Dimitriadis, Vasiliki Pappa
BACKGROUND: Myelodysplastic syndromes (MDS) are clonal hematopoietic stem cell disorders with unknown aetiology. Multiparameter flow cytometry (MFC) is being evaluated for the diagnosis and prognosis of MDS. METHODS: In the present study, five-color MFC was performed on bone marrow aspirates of 50 untreated patients, newly diagnosed with MDS and 27 age matched control samples. Patients were classified according to World Health Organization 2016, International Prognostic Scoring System (IPSS), and Revised IPSS (IPSS-R)...
October 17, 2018: Cytometry. Part B, Clinical Cytometry
Catarina Simões, Isabel Silva, Anabela Carvalho, Sandra Silva, Susana Santos, Gilberto Marques, André Ribeiro, Adriana Roque, José Carda, A B Sarmento-Ribeiro, Maria do Rosário Domingues, Letícia Ribeiro, Artur Paiva
BACKGROUND: Vδ1+ T cells, a subset of γδ T cells, are responsible for innate-like immune responses. Recently, an anti-tumor function mediated by MHC-unrestricted recognition of lipid and stress molecules, has also been described in these cells. This study aimed to quantify and phenotypically characterize circulating Vδ1+ T cells in B cell Chronic Lymphocytic Leukemia (CLL) and Monoclonal B cell lymphocytosis (MBL). METHODS: This study enrolled 58 individuals distributed in five groups: Binet B and C CLL (n = 9), Binet A CLL (n = 26), High count-MBL (n = 10), Low count-MBL (n = 5), and a control group (n = 8)...
October 17, 2018: Cytometry. Part B, Clinical Cytometry
John F Marcelletti, Branimir I Sikic, Larry D Cripe, Elisabeth Paietta
BACKGROUND: Multidrug resistance (MDR) transporter proteins such as P-glycoprotein (P-gp) efflux a variety of chemotherapeutic drugs from acute myeloid leukemia (AML) blasts leading to clinical drug resistance. METHODS: This study examined heterogeneity of MDR functional efflux by AML blasts using two flow cytometry bioassays. Bone marrow specimens (N = 50) from elderly patients with newly diagnosed AML were analyzed for CD34+ blasts with MDR efflux function. Efflux was measured with a fluorescent dye (DiOC2 ) as a surrogate for oncology drugs that are substrates for MDR efflux...
October 17, 2018: Cytometry. Part B, Clinical Cytometry
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