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Immunophenotyping by flow cytometry

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https://www.readbyqxmd.com/read/28067893/vemurafenib-and-trametinib-reduce-expression-of-ctgf-and-il-8-in-v600e-braf-melanoma-cells
#1
Mariusz L Hartman, Michal Rozanski, Marta Osrodek, Izabela Zalesna, Malgorzata Czyz
Clinical evidence has revealed that while RAS/RAF/MEK/ERK pathway is a crucial component of melanomagenesis, other signaling pathways can also contribute to the malignant growth and development of resistance to targeted therapies. We explored the response of (V600E)BRAF melanoma cells derived from surgical specimens and grown in stem cell medium to vemurafenib and trametinib, drugs targeting the activity of (V600E)BRAF and MEK1/2, respectively. Cell growth and apoptosis were monitored by real-time imaging system, immunophenotype and cell cycle by flow cytometry, gene expression by quantitative real-time PCR, immunoblotting and enzyme-linked immunosorbent assay...
January 9, 2017: Laboratory Investigation; a Journal of Technical Methods and Pathology
https://www.readbyqxmd.com/read/28063722/concordance-of-two-approaches-in-monitoring-of-minimal-residual-disease-in-b-precursor-acute-lymphoblastic-leukemia-fusion-transcripts-and-leukemia-associated-immunophenotypes
#2
Ying-Jung Huang, Elaine Coustan-Smith, Hsiao-Wen Kao, Hsi-Che Liu, Shih-Hsiang Chen, Chih-Cheng Hsiao, Chao-Ping Yang, Tang-Her Jaing, Ting-Chi Yeh, Ming-Chung Kuo, Chang-Liang Lai, Chia-Hui Chang, Dario Campana, Der-Cherng Liang, Lee-Yung Shih
BACKGROUND: Real-time quantitative polymerase chain reaction (RQ-PCR) for fusion transcripts and flow cytometry for leukemia-specific markers are widely used for minimal residual disease (MRD) detection in acute lymphoblastic leukemia, but the relation between the results of either method is unclear. METHODS: Mononucleated cells from 108 bone marrow samples collected from 55 B-precursor acute lymphoblastic leukemia patients (30 with t(12;21)/ETV6-RUNX1, 16 with t(9;22)/BCR-ABL1 and nine with t(1;19)/TCF3-PBX1) were examined in tandem by RQ-PCR and six-color flow cytometry...
January 4, 2017: Journal of the Formosan Medical Association, Taiwan Yi Zhi
https://www.readbyqxmd.com/read/28055240/multimodality-technologies-in-the-assessment-of-hematolymphoid-neoplasms
#3
Shiyong Li, David L Jaye, Kyle T Bradley, Linsheng Zhang, Debra Saxe, George Deeb, Charlie E Hill, Karen P Mann
Accurate assessment of tissues for hematolymphoid neoplasms requires an integrated multiparameter approach. Although morphologic examination by light microscopy remains the mainstay of initial assessment for hematolymphoid neoplasms, immunophenotypic analysis by immunohistochemistry and/or flow cytometry is essential to determine the pattern of differentiation and to detect minimal disease when morphology is inconclusive. In some cases, immunophenotypic analysis provides additional information for targeted immunotherapy and prognostication...
January 5, 2017: Archives of Pathology & Laboratory Medicine
https://www.readbyqxmd.com/read/28055237/enrichment-propagation-and-characterization-of-mouse-testis-derived-mesenchymal-stromal-cells
#4
Mai Ahmed, Myret Ghabriel, Asma Amleh
The therapeutic potential of multipotent stromal cells (MSCs) largely depends on the isolation and expansion methods used. In this study, we propose a laminin-based technique to select and enrich for MSCs isolated from the mouse testis. Primary cell cultures were prepared from juvenile mouse testes and the capacity to generate colony forming units together with population doubling time (PDT) during expansion were determined. The identity of MSCs was assayed using reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry for the active expression of cell surface markers, such as CD44, CD73, and CD29; absence of the CD45 hematopoietic cell marker; and in vitro differentiation of the cells into osteoblasts and adipocytes...
January 5, 2017: Cellular Reprogramming
https://www.readbyqxmd.com/read/28035134/upregulation-of-cc-chemokine-receptor-7-ccr7-enables-migration-of-xenogeneic-human-adipose-derived-mesenchymal-stem-cells-to-rat-secondary-lymphoid-organs
#5
Tian Ma, Shao-Liang Luan, Hong Huang, Xing-Kun Sun, Yan-Mei Yang, Hui Zhang, Wei-Dong Han, Hong Li, Yan Han
BACKGROUND CC chemokine receptor 7 (CCR7) expression is vital for cell migration to secondary lymphoid organs (SLOs). Our previous work showed that inducing CCR7 expression enabled syngeneic mesenchymal stem cells (MSCs) to migrate into SLOs, resulting in enhanced immunosuppressive performance in mice. Given that human adipose-derived stem cells (hASCs) are widely used in clinical therapy, we further investigated whether upregulation of CCR7 enables xenogeneic hASCs to migrate to rat SLOs. MATERIAL AND METHODS hASCs rarely express CCR7; therefore, hASCs were transfected with lentivirus encoding rat CCR7 (rCCR7) plus green fluorescence protein (GFP) or GFP alone...
December 30, 2016: Medical Science Monitor: International Medical Journal of Experimental and Clinical Research
https://www.readbyqxmd.com/read/28024875/comparative-characterization-of-mesenchymal-stromal-cells-from-multiple-abdominal-adipose-tissues-and-enrichment-of-angiogenic-ability-via-cd146-molecule
#6
Na Eun Lee, Sung Joo Kim, Seung-Jib Yang, Sung-Yeon Joo, Hyojun Park, Kyo Won Lee, Heung-Mo Yang, Jae Berm Park
BACKGROUND: There are various types of adipose tissue in the human body, and their morphology is known to be closely related to cell function and metabolism. However, the functional differences among the mesenchymal stromal cells (MSCs) of different abdominal adipose tissues have not been clearly elucidated. METHODS: MSCs were isolated from different abdominal adipose tissues according to their regional distribution and included superficial subcutaneous, deep subcutaneous, omentum, mesentery and retroperitoneal MSCs...
February 2017: Cytotherapy
https://www.readbyqxmd.com/read/28024508/-reconstruction-of-humanized-bone-marrow-niche-in-immunodeficiency-mouse
#7
Huan Chen, Zheng Tian, Bin Li, Hai-Yan Xing, Huan Li, Ke-Jing Tang, Min Wang, Qing Rao
OBJECTIVE: To reconstruct a human bone marrow niche in immunodeficiency mouse (NOD/SCID) so as to provide a model for observing the effect of abnormal BM niche on the occurence and development of leukemia. METHODS: Human platelet lysate(HPL) was obtained by repeated freezing and thawing of concentrated platelet. Bone marrow-derived mesenchymal stem cells were cultured in α-minimal essential medium (α-MEM) containing 10% HPL or 10% FBS. The morphology, cell phenotype, multilineage differentiation potential in vitro and proliferation capacity between the mesenchymal stem cells cultured with HPL or FBS were compared...
December 2016: Zhongguo Shi Yan Xue Ye Xue za Zhi
https://www.readbyqxmd.com/read/28024467/-features-of-immunophenotypes-and-characteristics-of-molecular-biology-and-cellular-genetics-of-aml-patients-with-cd4-and-cd7-expression
#8
Tie-Qiang Liu, Shan Huang, Bo Yao, Zhi-Qing Liu, Chang-Lin Yu, Jian-Hui Qiao, Qi-Yun Sun, Kai-Xun Hu, Ya-Jing Huang, Rui Zhang, Yu-Fang Li, Juan Bai, Yu-Jing Sun, Bing-Xia Li, Dong-Mei Wang, Yi Wang, Mei Guo
OBJECTIVE: To explore the features of immunophenotypes and the characteristics of molecular biology and cellular genetics of AML patients with CD7 and CD4 expression. METHODS: The immunophenotypical markers of AML cells were detected by multiple parameter flow cytometry; the expression of WT1, MDK, ETO, PML-RaRa and BCR-ABL were detected by RT-PCR; and cellular features were analyzed by R-band in 304 patients. The patients were divided into three groups according to their immunophenotypes: AML with CD7 expression (CD7 group), AML with CD4 expression(CD4 group) and AML without CD7 and CD4 expression (common AML group)...
December 2016: Zhongguo Shi Yan Xue Ye Xue za Zhi
https://www.readbyqxmd.com/read/28007666/real-time-immunophenotyping-of-leukocyte-subsets-early-after-double-cord-blood-transplant-predicts-graft-function
#9
Jianqiang Li, Ian Nicoud, Joseph Blake, David Oliver, Emily Cox, Shelly Heimfeld, Filippo Milano, Suzan Imren, Colleen Delaney
Cord blood transplant (CBT) recipients are at increased risk for delayed engraftment and primary graft failure (PGF), complications that are often indistinguishable early post-transplant. Current assays fail to accurately identify recipients with slow hematopoietic recovery and distinguish them from those with pending graft failure. To address this, we prospectively examined the kinetics of immune cell subset recovery in the peripheral blood of 39 patients on days 7 and 14 after double-unit CBT (dCBT) by multiparametric flow cytometry analysis, which we called real time immunophenotyping (RTIP)...
December 19, 2016: Biology of Blood and Marrow Transplantation
https://www.readbyqxmd.com/read/28004597/functional-proteomic-insights-in-b-cell-chronic-lymphocytic-leukemia
#10
Paula Díez, Rafael Góngora, Alberto Orfao, Manuel Fuentes
B cell chronic lymphocytic leukemia (B-CLL) is a hematological malignancy considered as the most common leukemia in the Western world. The understanding of B cell differentiation is crucial for the diagnosis, prognosis, and treatment of the disease. Areas covered: In this review, B-cell ontogeny and its relation with the CLL development, in combination with the proteomic approaches which could provide a deep characterization of the disease through the characterization of the cellular signaling pathways involved in the pathological cells is described...
December 30, 2016: Expert Review of Proteomics
https://www.readbyqxmd.com/read/28003262/defining-the-clonality-of-peripheral-t-cell-lymphomas-using-rna-seq
#11
Scott D Brown, Greg Hapgood, Christian Steidl, Andrew P Weng, Kerry J Savage, Robert A Holt
MOTIVATION: In T-cell lymphoma, malignant T cells arising from a founding clone share an identical T cell receptor (TCR) and can be identified by the over-representation of this TCR relative to TCRs from the patient's repertoire of normal T cells. Here, we demonstrate that TCR information extracted from RNA-seq data can provide a higher resolution view of peripheral T cell lymphomas (PTCLs) than that provided by conventional methods. RESULTS: For 60 subjects with PTCL, flow cytometry/FACS was used to identify and sort aberrant T cell populations from diagnostic lymph node cell suspensions...
December 21, 2016: Bioinformatics
https://www.readbyqxmd.com/read/28003154/antipsychotics-influence-toll-like-receptor-tlr-expression-and-its-relationship-with-cognitive-functions-in-schizophrenia
#12
Szabolcs Kéri, Csilla Szabó, Oguz Kelemen
Increasing evidence suggests that altered immune functions are related to the pathophysiology of schizophrenia. Relatively little information is available on Toll-like receptors (TLRs), which are implicated in the recognition of molecular patterns associated with pathogens and internal cellular damage signals. By using immunophenotyping and flow cytometry, we investigated TLRs in CD14+ monocytes, CD4+CD25+Foxp3+ regulatory T cells (Treg), and CD3+CD4+CD25+ activated T cells (Tact) in 35 drug-naïve patients with schizophrenia before and after an 8-week period of antipsychotic treatment with risperidone or olanzapine...
December 19, 2016: Brain, Behavior, and Immunity
https://www.readbyqxmd.com/read/27998726/characterization-and-purification-of-neoplastic-cells-of-nodular-lymphocyte-predominant-hodgkin-lymphoma-from-lymph-nodes-by-flow-cytometry-and-flow-cytometric-cell-sorting
#13
Jonathan R Fromm, Anju Thomas, Brent L Wood
We report the flow cytometric (FC) identification and characterization of lymphocyte predominant (LP) cells from lymph nodes involved by nodular lymphocyte predominant Hodgkin lymphoma (NLPHL). First, we immunophenotyped the NLPHL cell line (DEV) confirming a germinal center immunophenotype, lack of expression of CD32 and CD58, and expression of CD54. Nineteen of 26 lymph nodes involved by NLPHL demonstrated a population with an LP immunophenotype (73%), which included expression of germinal center markers (CD75/Bcl-6-positive, CD32-weak/negative without CD10), a B-cell immunophenotype (CD19/CD20/CD40(+)), IgD and/or IgM expression (67%), and lack of programmed death-ligand 1/ligand 2...
December 17, 2016: American Journal of Pathology
https://www.readbyqxmd.com/read/27981867/phenotypic-and-functional-characterization-of-a-bortezomib-resistant-multiple-myeloma-cell-line-by-flow-and-mass-cytometry
#14
Linda B Baughn, Zohar Sachs, Klara E Noble-Orcutt, Amit Mitra, Brian G Van Ness, Michael A Linden
Multiple myeloma (MM) is an incurable malignant plasma cell neoplasm. Proteasome inhibitors including Bortezomib (Bz) are used to treat MM, and treatment failure due to drug resistance occurs. Bz-sensitive and -resistant MM cells have distinct immunophenotypic signatures that correlate with clinical outcome. These changes can be identified by fluorescence-based cytometry (FBC), however, FBC is rarely used in predicting Bz resistance. Mass cytometry (MC) is a recently developed variation of flow cytometry that detects heavy metal-ion tagged antibodies using time-of-flight mass spectrometry allowing for detection of up to 38 epitopes simultaneously in a single cell, without significant overlap, exceeding the dimensionality of FBC 3-4-fold...
December 16, 2016: Leukemia & Lymphoma
https://www.readbyqxmd.com/read/27942026/characterisation-of-the-immunophenotype-of-dogs-with-primary-immune-mediated-haemolytic-anaemia
#15
James W Swann, Kelly Woods, Ying Wu, Barbara Glanemann, Oliver A Garden
BACKGROUND: Immune-mediated haemolytic anaemia (IMHA) is reported to be the most common autoimmune disease of dogs, resulting in significant morbidity and mortality in affected animals. Haemolysis is caused by the action of autoantibodies, but the immunological changes that result in their production have not been elucidated. AIMS: To investigate the frequency of regulatory T cells (Tregs) and other lymphocyte subsets and to measure serum concentrations of cytokines and peripheral blood mononuclear cell expression of cytokine genes in dogs with IMHA, healthy dogs and dogs with inflammatory diseases...
2016: PloS One
https://www.readbyqxmd.com/read/27907250/the-transcriptional-co-activator-bob1-is-associated-with-pathologic-b-cell-responses-in-autoimmune-tissue-inflammation
#16
Maria J Levels, Melissa N Van Tok, Tineke Cantaert, Juan D Cañete, Frans G M Kroese, Kristine Germar, Hergen Spits, Dominique L P Baeten, Nataliya G Yeremenko
OBJECTIVE: The molecular mechanisms steering abnormal B cell responses in autoimmune diseases remain poorly understood. In this study we aimed to identify molecular switches controlling pathological B cell responses in rheumatoid arthritis (RA). METHODS: Candidate molecules were identified by gene expression profiling of RA synovitis and validated by qPCR and immunohistochemistry. B cell-specific expression was confirmed by immunofluorescence, immunoblotting and flow cytometry...
December 1, 2016: Arthritis & Rheumatology
https://www.readbyqxmd.com/read/27906060/discrete-adipose-derived-stem-cell-subpopulations-may-display-differential-functionality-after-in-vitro-expansion-despite-convergence-to-a-common-phenotype-distribution
#17
Frederik Mølgaard Nielsen, Simone Elkjær Riis, Jens Isak Andersen, Raphaëlle Lesage, Trine Fink, Cristian Pablo Pennisi, Vladimir Zachar
BACKGROUND: Complex immunophenotypic repertoires defining discrete adipose-derived stem cell (ASC) subpopulations may hold a key toward identifying predictors of clinical utility. To this end, we sorted out of the freshly established ASCs four subpopulations (SPs) according to a specific pattern of co-expression of six surface markers, the CD34, CD73, CD90, CD105, CD146, and CD271, using polychromatic flow cytometry. METHOD: Using flow cytometry-associated cell sorting and analysis, gating parameters were set to select for a CD73(+)CD90(+)CD105(+) phenotype plus one of the four following combinations, CD34(-)CD146(-)CD271(-) (SP1), CD34(-)CD146(+)CD271(-) (SP2), CD34(+)CD146(+)CD271(-) (SP3), and CD34(-)CD146(+)CD271(+) (SP4)...
December 1, 2016: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/27902915/characterization-of-chicken-splenic-derived-dendritic-cells-following-vaccine-and-very-virulent-strains-of-infectious-bursal-disease-virus-infection
#18
A R Yasmin, S K Yeap, M Hair-Bejo, A R Omar
Studies have shown that infectious bursal disease virus (IBDV) infects lymphoid cells, mainly B cells and macrophages. This study was aimed to examine the involvement of chicken splenic-derived dendritic cells (ch-sDCs) in specific-pathogen-free chickens following inoculation with IBDV vaccine strain (D78) and a very virulent (vv) strain (UPM0081). Following IBDV infection, enriched activated ch-sDCs were collected by using the negative selection method and were examined based on morphology and immunophenotyping to confirm the isolation method for dendritic cells (DCs)...
December 2016: Avian Diseases
https://www.readbyqxmd.com/read/27899414/sentinel-case-of-richter-transformation-from-chronic-lymphocytic-leukaemia-small-lymphocytic-lymphoma-to-cd3-diffuse-large-b-cell-lymphoma
#19
Ali Ismail, Jawed A Mallick, Dahui Qin, Mohammad O Hussaini
AIM: To report the first case of a Richter syndrome where small lymphocytic lymphoma (SLL) progressed to a CD3+ diffuse large B-cell lymphoma (DLBCL). METHODS: Macrodissection of small and large cell lymphomatous components was performed. This was followed by flow cytometric analysis along with molecular B-cell immunoglobulin (heavy and light chains) and T-cell receptor (γ and β chains) gene rearrangement studies to investigate a clonal relationship between the components...
November 29, 2016: Journal of Clinical Pathology
https://www.readbyqxmd.com/read/27899238/goat-umbilical-cord-cells-are-permissive-to-small-ruminant-lentivirus-infection-in-vitro
#20
Gabrielle R Martins, Rebeca C Marinho, Rosivaldo Q Bezerra Junior, Antoniel de O Alves, Lilia M C Câmara, Luiz C Albuquerque-Pinto, Maria F da S Teixeira
Small ruminant lentiviruses isolated from peripheral blood leukocytes and target organs can be propagated in vitro in fibroblasts derived from goat synovial membrane cells. These cells are obtained from tissues collected from embryos or fetuses and are necessary for the establishment of the fibroblast primary culture. A new alternative type of host cells, derived from goat umbilical cord, was isolated and characterized phenotypically with its main purpose being to obtain cell monolayers that could be used for the diagnosis and isolation of small ruminant lentiviruses in cell culture...
November 19, 2016: Brazilian Journal of Microbiology: [publication of the Brazilian Society for Microbiology]
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