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flow cytometry immunophenotyping

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https://www.readbyqxmd.com/read/28927728/cryopreservation-of-human-whole-blood-allows-immunophenotyping-by-flow-cytometry-up-to-30days-after-cell-isolation
#1
R Madelaine Paredes, Douglas K Tadaki, Amanda Sooter, Fabia Gamboni, C D R Forest Sheppard
Immunophenotyping of whole blood (WB) by flow cytometry (FC) is used clinically to assess a patient's immune status and also in biomedical research. Current protocols recommend storage of immunolabeled samples at 4°C with FC analysis to be completed within seven days. This data acquisition window can be extended to up to one year post-labeling, but this requires cryopreservation of the samples at ultra-low temperatures (≤-80°C or in liquid nitrogen). In this study we optimized a standardized cryopreservation protocol to enable preservation of immunolabeled, human WB samples at -20°C for FC and tested its effectiveness after 0, 5, 15 or 30days...
September 16, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28925935/technical-advances-in-the-measurement-of-residual-disease-in-acute-myeloid-leukemia
#2
REVIEW
Gregory W Roloff, Catherine Lai, Christopher S Hourigan, Laura W Dillon
Outcomes for those diagnosed with acute myeloid leukemia (AML) remain poor. It has been widely established that persistent residual leukemic burden, often referred to as measurable or minimal residual disease (MRD), after induction therapy or at the time of hematopoietic stem cell transplant (HSCT) is highly predictive for adverse clinical outcomes and can be used to identify patients likely to experience clinically evident relapse. As a result of inherent genetic and molecular heterogeneity in AML, there is no uniform method or protocol for MRD measurement to encompass all cases...
September 19, 2017: Journal of Clinical Medicine
https://www.readbyqxmd.com/read/28916595/normative-data-for-flow-cytometry-immunophenotyping-of-benign-lymph-nodes-sampled-by-surgical-biopsy
#3
Gregory David Scott, Susan K Atwater, Dita A Gratzinger
AIMS: To create clinically relevant normative flow cytometry data for understudied benign lymph nodes and characterise outliers. METHODS: Clinical, histological and flow cytometry data were collected and distributions summarised for 380 benign lymph node excisional biopsies. Outliers for kappa:lambda light chain ratio, CD10:CD19 coexpression, CD5:CD19 coexpression, CD4:CD8 ratios and CD7 loss were summarised for histological pattern, concomitant diseases and follow-up course...
September 15, 2017: Journal of Clinical Pathology
https://www.readbyqxmd.com/read/28888074/early-recovery-of-circulating-immature-b-cells-in-b-lymphoblastic-leukemia-patients-after-cd19-targeted-car-t-cell-therapy-a-pitfall-for-minimal-residual-disease-detection
#4
Wenbin Xiao, Dalia Salem, Catherine McCoy, Daniel Lee, Nirali N Shah, Maryalice Stetler-Stevenson, Constance M Yuan
BACKGROUND: CD19-targeted chimeric-antigen receptor-modified T-cells (CAR-T) are promising in the treatment of refractory B-lymphoblastic leukemia (B-ALL). Minimal residual disease (MRD) detection by multicolor flow cytometry (FCM) is critical to distinguish B-ALL MRD from regenerating, non-neoplastic B-cell populations. METHODS: FCM was performed on samples from 9 patients with B-ALL treated with CAR-T. RESULTS: All 9 patients showed response to CAR-T...
September 9, 2017: Cytometry. Part B, Clinical Cytometry
https://www.readbyqxmd.com/read/28883278/loss-of-cd45-expression-at-relapse-of-acute-myeloid-leukemia
#5
Katsuya Yamamoto, Kimikazu Yakushijin, Keiji Kurata, Yu Mizutani, Yumiko Inui, Kiyoaki Uryu, Shinichiro Kawamoto, Takeshi Sugimoto, Hiroshi Matsuoka, Hironobu Minami
A 49-year-old female was initially diagnosed with acute myeloid leukemia (AML) M4 with a CD45+CD13+CD33+CD34-HLA-DR+ immunophenotype. She underwent allogeneic bone marrow transplantation, but the disease recurred. The bone marrow was infiltrated with 87.0% blasts negative for myeloperoxidase (MPO) staining. Immunophenotyping by flow cytometry identified the presence of a CD45-negative blast population. These blasts exhibited a CD13+CD33+CD19-CD10-CD34-HLA-DR- immunophenotype. The lack of CD45 expression is often observed in B-cell acute lymphoblastic leukemia, whereas CD45-negative AML is extremely rare; only one older male with AML-M0 has been reported...
2017: [Rinshō Ketsueki] the Japanese Journal of Clinical Hematology
https://www.readbyqxmd.com/read/28877231/functional-characterization-of-t-cells-from-palatine-tonsils-in-patients-with-chronic-tonsillitis
#6
Katharina Geißler, Robby Markwart, Robert Pascal Requardt, Cynthia Weigel, Katja Schubert, André Scherag, Ignacio Rubio, Orlando Guntinas-Lichius
The palatine tonsils, localized in the oropharynx, are easily accessible secondary lymphoid tissue in humans. Inflammation of the palatine tonsils, local and chronic in case of chronic tonsillitis (CT) or acute in the presence of a peritonsillar abscess (PTA), ranks among the most common diseases in otolaryngology. However, the functionality of tonsillar immune cells, notably T-cells, in the context of these immune pathologies is poorly understood. We have examined the functional status of human tonsillar T-cells in CT and compared it to the acute inflammatory setting of a PTA...
2017: PloS One
https://www.readbyqxmd.com/read/28873356/follicular-lymphoma-with-hyaline-vascular-castleman-like-features-analysis-of-6-cases-and-review-of-the-literature
#7
Sergio Pina-Oviedo, Roberto N Miranda, Pei Lin, John T Manning, L Jeffrey Medeiros
Follicular lymphoma (FL) with features reminiscent of hyaline-vascular Castleman disease (CD) is an unusual morphologic variant that may create diagnostic difficulties. To our knowledge, only 5 cases of this variant have been reported. We describe the clinicopathologic features of 6 cases including 2 men and 4 women with a median age of 63years (range, 41-77). Morphologically, all lymph node biopsy specimens showed at least a focal area of conventional FL; 4 cases showed neoplastic follicles with hyalinized blood vessels penetrating into germinal centers (lollipop-like lesions); 4 cases had interfollicular areas with increased vascular stroma, 2 cases showed small neoplastic follicles with prominent, onion skin-like mantle zones, and 1 case showed two or more germinal centers within follicles (twinning)...
September 2, 2017: Human Pathology
https://www.readbyqxmd.com/read/28870198/up-modulation-of-plc-%C3%AE-2-reduces-the-number-and-malignancy-of-triple-negative-breast-tumor-cells-with-a-cd133-epcam-phenotype-a-promising-target-for-preventing-progression-of-tnbc
#8
Federica Brugnoli, Silvia Grassilli, Paola Lanuti, Marco Marchisio, Yasamin Al-Qassab, Federica Vezzali, Silvano Capitani, Valeria Bertagnolo
BACKGROUND: The malignant potential of triple negative breast cancer (TNBC) is also dependent on a sub-population of cells with a stem-like phenotype. Among the cancer stem cell markers, CD133 and EpCAM strongly correlate with breast tumor aggressiveness, suggesting that simultaneous targeting of the two surface antigens may be beneficial in treatment of TNBC. Since in TNBC-derived cells we demonstrated that PLC-β2 induces the conversion of CD133(high) to CD133(low) cells, here we explored its possible role in down-modulating the expression of both CD133 and EpCAM and, ultimately, in reducing the number of TNBC cells with a stem-like phenotype...
September 4, 2017: BMC Cancer
https://www.readbyqxmd.com/read/28862348/immunophenotype-of-peripheral-blood-lymphocytes-in-dogs-with-inflammatory-bowel-disease
#9
A Galler, B C Rütgen, E Haas, A Saalmüller, R A Hirt, W Gerner, I Schwendenwein, B Richter, J G Thalhammer, N Luckschander-Zeller
BACKGROUND: Inflammatory bowel disease (IBD) is common in dogs. Despite the known importance of intestinal lymphocytes in its pathogenesis, little is known about the role of peripheral blood lymphocytes (PBLs) in IBD. OBJECTIVES: The aims of this study were (1) comparison of PBLs analyzed by flow cytometry (FCM) in IBD dogs and healthy controls and (2) comparison of PBLs in IBD dogs at the time of diagnosis and in dogs in clinical remission. ANIMALS: Whole blood samples of 19 IBD dogs at the time of diagnosis and blood samples of 6 dogs in clinical remission were collected...
September 1, 2017: Journal of Veterinary Internal Medicine
https://www.readbyqxmd.com/read/28844128/a-simple-method-to-isolate-and-expand-human-umbilical-cord-derived-mesenchymal-stem-cells-using-explant-method-and-umbilical-cord-blood-serum
#10
Ghmkin Hassan, Issam Kasem, Chadi Soukkarieh, Majd Aljamali
Background and Objectives: Mesenchymal stem cells (MSCs) are multipotent stem cells that can be isolated from umbilical cords and are therapeutically used because of their ability to differentiate into various types of cells, in addition to their immunosuppressive and anti-inflammatory properties. Fetal bovine serum (FBS), considered as the standard additive when isolating and culturing MSCs, has a major limitation related to its animal origin. Here, we employed a simple and economically efficient protocol to isolate MSCs from human umbilical cord tissues without using digestive enzymes and replacing FBS with umbilical cord blood serum (CBS)...
August 31, 2017: International Journal of Stem Cells
https://www.readbyqxmd.com/read/28830803/acute-promyelocytic-leukemia-in-a-patient-with-chronic-lymphocytic-leukemia-a-case-report
#11
Prajwal Boddu, Ellen Schlette, Beenu Thakral, Guillin Tang, Naveen Pemmaraju, Tapan Kadia, Alessandra Ferrajoli, Farhad Ravandi, William Wierda, Nitin Jain
Chronic lymphocytic leukemia (CLL) is known to be associated rarely with myeloid malignancies such as acute myelogenous leukemia. In this article, we report an extremely rare occurrence of acute promyelocytic leukemia in a patient with CLL. A 71-year-old man first presented to our clinic with a diagnosis of CLL and worsening motor neuropathy symptoms. It was suspected that his CLL might be contributing to the neuropathy as a paraneoplastic syndrome, and he was treated with rituximab monotherapy in weekly doses for the 1st month and monthly treatments thereafter...
August 16, 2017: Hematology/oncology and Stem Cell Therapy
https://www.readbyqxmd.com/read/28815514/intraepithelial-lymphocyte-immunophenotype-a-useful-tool-in-the-diagnosis-of-celiac-disease
#12
Rebeca Saborido, Nazareth Martinón, Alexandra Regueiro, Vanesa Crujeiras, Pablo Eiras, Rosaura Leis
According to new ESPGHAN guidelines, gluten challenge is considered necessary when there is doubt about the initial diagnosis of celiac disease (CD). The main aim of this study was to quantify intraepithelial lymphocyte (IEL) immunophenotype on celiac patients on gluten-containing diet (GCD) compared to those on gluten-free diet (GFD). Another aim was to evaluate the clinical utility of IELs in the CD diagnosis, especially in selected patients on GFD where diagnostic uncertainty remains. IEL immunophenotype (TCRγδ and NK-like IELs) were studied by flow cytometry in 111 children with CD (81 children with CD on GCD and 30 celiac patients on GFD) and a control group (10 children)...
August 16, 2017: Journal of Physiology and Biochemistry
https://www.readbyqxmd.com/read/28800175/mesenchymal-stromal-cells-contribute-to-quiescence-of-therapy-resistant-leukemic-cells-in-acute-myeloid-leukemia
#13
Wenwen Wang, Tilmann Bochtler, Patrick Wuchter, Linda Manta, Haiju He, Volker Eckstein, Anthony D Ho, Christoph Lutz
OBJECTIVE: Persistence of leukemic cells after induction therapy has been shown to correlate with poor survival in acute myeloid leukemia (AML). In this study we tested if human mesenchymal stromal cells (hMSC) have protective effects on leukemic cells undergoing chemotherapy. METHODS: Persistent disease was used as marker to identify cases with therapy resistant leukemic cells in 95 AML patients. Immunophenotyping, cell cycle and apoptosis assays were assessed by flow-cytometry...
August 11, 2017: European Journal of Haematology
https://www.readbyqxmd.com/read/28792255/host-related-immunodeficiency-in-the-development-of-multiple-myeloma
#14
Talib Dosani, Sham Mailankody, Neha Korde, Elisabet Manasanch, Manisha Bhutani, Nishant Tageja, Mark Roschewski, Mary Kwok, Dickran Kazandjian, Rene Costello, Debra Burton, Yong Zhang, David Liewehr, Seth M Steinberg, Irina Maric, Ola Landgren
Host-related immunodeficiency is known to play a role in the development of multiple myeloma (MM) from its precursor conditions (monoclonal gammopathy of undetermined significance, MGUS, smoldering multiple myeloma, SMM). In order to understand the underlying immune changes in this process, we characterized immune patterns from MGUS to SMM to MM. We further sought to identify potential novel immune biomarkers that may predict progression of SMM to MM. We characterized patterns of circulating lymphocytes in 181 patients using multiparametric flow cytometry...
August 9, 2017: Leukemia & Lymphoma
https://www.readbyqxmd.com/read/28792162/immunological-subtypes-of-acute-lymphoblastic-leukemia-beyond-morphology-experience-from-kidwai-state-cancer-institute-bengaluru-india
#15
Namrata N Rajkumar, Raghavendra H Vijay
INTRODUCTION: Acute lymphoblastic leukemia (ALL) is disease of lymphoid precursors and is the most common cancer. Diagnosis of ALL is made by evaluating morphology and flowcytometric Immunophenotyping (FCI)and is an important adjunct in diagnosis and determining treatment in ALL, with availability of extensive monoclonal antibodies in the recent years there is tremendous progress in the field of FCI, and is a requirement by World Health Organisation for the classification of acute lymphoblastic Leukemia...
July 2017: Journal of the Association of Physicians of India
https://www.readbyqxmd.com/read/28789968/b-cell-phenotypes-signaling-and-their-roles-in-secretion-of-antibodies-in-systemic-lupus-erythematosus
#16
Yoshiya Tanaka, Satoshi Kubo, Shigeru Iwata, Maiko Yoshikawa, Shingo Nakayamada
B cells play a pivotal role in the initiation and perpetuation of SLE. Because SLE is molecularly and clinically heterogeneous, efficacious targeted therapy to clinical remission has not yet been established in SLE. We have found i) statistical clustering between Tfh cells and class-switched memory B cells and the upregulated transition from CXCR5(+) IgM memory B cells to CXCR3(+) class-switched memory B cells in SLE by 8-color flow cytometry, ii) the involvement of Syk, Btk and JAK in the activation and differentiation of B cells in SLE, iii) SLE patients was divided to 3 groups based on immunophenotypic analysis and statistical analysis and patients in the Tfh/class-switched B cell-dominant group were most refractory to conventional therapies although 3 groups had similar clinical features...
August 5, 2017: Clinical Immunology: the Official Journal of the Clinical Immunology Society
https://www.readbyqxmd.com/read/28777444/intracellular-cytokine-detection-by-flow-cytometry-in-surface-marker-defined-human-peripheral-blood-mononuclear-t-cells
#17
Fredine T Lauer, Jesse L Denson, Ellen Beswick, Scott W Burchiel
In a recent unit in this series, protocols for the isolation, cryopreservation, thawing, and immunophenotyping of HPBMC isolated from peripheral whole blood using cell surface marker (CSM) staining and multi-color flow cytometry analysis were presented. The current procedure describes the detection and quantification of CSM and intracellular markers (ICM), including transcription factors and cytokines, following activation and differentiation of CD4+ T-cells using multi-color flow cytometry. Results indicated that repeatable and robust detection of ICM could be obtained in surface marker-defined T cells that identify functional subsets of cells...
August 4, 2017: Current Protocols in Toxicology
https://www.readbyqxmd.com/read/28766537/-multiple-myeloma-maintenance-therapy-after-autologous-hematopoietic-stem-cell-transplantation-depending-on-minimal-residual-disease
#18
M V Solovyev, L P Mendeleeva, O S Pokrovskaya, M V Nareyko, M V Firsova, I V Galtseva, Yu O Davydova, N M Kapranov, L A Kuzmina, E G Gemdzhian, V G Savchenko
AIM: To determine the efficiency of maintenance therapy with bortezomib in patients with multiple myeloma (MM) who have achieved complete remission (CR) after autologous hematopoietic stem cell (auto-HSCT), depending on the presence of minimal residual disease (MRD). SUBJECTS AND METHODS: In January 2014 to February 2016, fifty-two MM patients (19 men and 33 women) aged 24 to 66 years (median 54 years), who had achieved CR after auto-HSCT, were randomized to perform maintenance therapy with bortezomib during a year...
2017: Terapevticheskiĭ Arkhiv
https://www.readbyqxmd.com/read/28764802/mesenchymal-stem-cells-derived-from-inflamed-dental-pulpal-and-gingival-tissue-a-potential-application-for-bone-formation
#19
Laura Tomasello, Rodolfo Mauceri, Antonina Coppola, Maria Pitrone, Giuseppe Pizzo, Giuseppina Campisi, Giuseppe Pizzolanti, Carla Giordano
BACKGROUND: Chronic periodontal disease is an infectious disease consisting of prolonged inflammation of the supporting tooth tissue and resulting in bone loss. Guided bone regeneration procedures have become common and safe treatments in dentistry, and in this context dental stem cells would represent the ideal solution as autologous cells. In this study, we verified the ability of dental pulp mesenchymal stem cells (DPSCs) and gingival mesenchymal stem cells (GMSCs) harvested from periodontally affected teeth to produce new mineralized bone tissue in vitro, and compared this to cells from healthy teeth...
August 1, 2017: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/28760670/standardization-of-8-color-flow-cytometry-across-different-flow-cytometer-instruments-a-feasibility-study-in-clinical-laboratories-in-switzerland
#20
Glier Hana, Heijnen Ingmar, Hauwel Mathieu, Dirks Jan, Quarroz Stéphane, Lehmann Thomas, Rovo Alicia, Arn Kornelius, Matthes Thomas, Hogan Cassandra, Keller Peter, Dudkiewicz Ewa, Stüssi Georg, Fernandez Paula
The EuroFlow Consortium developed a fully standardized flow cytometric approach from instrument settings, through antibody panel, reagents and sample preparation protocols, to data acquisition and analysis. The Swiss Cytometry Society (SCS) promoted a study to evaluate the feasibility of using such standardized measurements of 8-color data across two different flow cytometry platforms - Becton Dickinson (BD) FACSCanto II and Beckman Coulter (BC) Navios, aiming at increasing reproducibility and inter-laboratory comparability of immunophenotypic data in clinical laboratories in Switzerland...
July 28, 2017: Journal of Immunological Methods
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