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Multicolor Flow Cytometry

Kieu-Suong Le, Patricia Amé-Thomas, Karin Tarte, Françoise Gondois-Rey, Samuel Granjeaud, Florence Orlanducci, Etienne D Foucher, Florence Broussais, Reda Bouabdallah, Thierry Fest, Dominique Leroux, Sapna Yadavilli, Patrick A Mayes, Luc Xerri, Daniel Olive
A better characterization of T-cell subsets in the microenvironment of classical Hodgkin lymphoma (cHL) would help to develop immunotherapies. Using multicolor flow cytometry, we identified in 6 of 43 cHL tissue samples a previously unrecognized subset of CD8 T cells coexpressing CXCR5 and inducible T-cell costimulator (ICOS) molecules (CD8CXCR5+ICOS+ ). These cells shared phenotypic features with follicular helper T (TFH ) cells including low CCR7 expression together with high expression of B-cell lymphoma-6, programmed cell death 1, B and T lymphocyte attenuator, CD200, and OX40...
August 14, 2018: Blood Advances
Nikunj Gevariya, Marjorie Besançon, Karine Robitaille, Valérie Picard, Lamoussa Diabaté, Anwar Alesawi, Pierre Julien, Yves Fradet, Alain Bergeron, Vincent Fradet
BACKGROUND: Several lines of evidence suggest effects of dietary fat on prostate cancer (PCa) development and progression. Targeting omega (ω)-3:ω6 fatty acids (FA) ratio could be beneficial against PCa by favorably modulating inflammation. Here, we studied the effects of ω3- and ω6-enriched diets on prostate tumor growth and inflammatory response in androgen-deprived and non-deprived conditions. METHODS: Immune-competent eugonadal and castrated C57BL/6 mice were injected with TRAMP-C2 prostate tumor cells and daily fed with ω3- or ω6-enriched diet...
August 2, 2018: Prostate
Aras Toker, Linh T Nguyen, Simone C Stone, Cindy Yang, Sarah R Katz, Patricia A Shaw, Blaise A Clarke, Danny A Ghazarian, Ayman S Al Habeeb, Alexandra M Easson, Wey Leong, David McCready, Michael Reedijk, Cynthia J Guidos, Trevor J Pugh, Marcus Q Bernardini, Pamela S Ohashi
PURPOSE: Regulatory T (Treg) cells expressing the transcription factor FOXP3 are essential for the maintenance of immunological self-tolerance but play a detrimental role in most cancers due to their ability to suppress antitumor immunity. The phenotype of human circulating Treg cells has been extensively studied, but less is known about tumor-infiltrating Treg cells. We studied the phenotype and function of tumor-infiltrating Treg cells in ovarian cancer and melanoma to identify potential Treg cell-associated molecules that can be targeted by tumor immunotherapies...
July 31, 2018: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
Andrzej Eljaszewicz, Lukasz Bolkun, Kamil Grubczak, Malgorzata Rusak, Tomasz Wasiluk, Milena Dabrowska, Piotr Radziwon, Wojciech Marlicz, Karol Kamiński, Janusz Kloczko, Marcin Moniuszko
Background: Acute lymphoblastic leukemia (ALL) is a malignant disease of lymphoid progenitor cells. ALL chemotherapy is associated with numerous side effects including neutropenia that is routinely prevented by the administration of growth factors such as granulocyte colony-stimulating factor (G-CSF). To date, the effects of G-CSF treatment on the level of mobilization of different stem and progenitor cells in ALL patients subjected to clinically effective chemotherapy have not been fully elucidated...
2018: Stem Cells International
Rita Folcarelli, Selma van Staveren, Roel Bouman, Bart Hilvering, Gerjen H Tinnevelt, Geert Postma, Oscar F van den Brink, Lutgarde M C Buydens, Nienke Vrisekoop, Leo Koenderman, Jeroen J Jansen
Multicolor Flow Cytometry (MFC)-based gating allows the selection of cellular (pheno)types based on their unique marker expression. Current manual gating practice is highly subjective and may remove relevant information to preclude discovery of cell populations with specific co-expression of multiple markers. Only multivariate approaches can extract such aspects of cell variability from multi-dimensional MFC data. We describe the novel method ECLIPSE (Elimination of Cells Lying in Patterns Similar to Endogeneity) to identify and characterize aberrant cells present in individuals out of homeostasis...
July 19, 2018: Scientific Reports
Kimberly A Muczynski, Nicolae Leca, Arthur E Anderson, Niamh Kieran, Susan K Anderson
Introduction: Current processing of renal biopsy samples provides limited information about immune mechanisms causing kidney injury and disease activity. We used flow cytometry with transplanted kidney biopsy samples to provide more information on the immune status of the kidney. Methods: To enhance the information available from a biopsy, we developed a technique for reducing a fraction of a renal biopsy sample to single cells for multicolor flow cytometry and quantitation of secreted cytokines present within the biopsy sample...
July 2018: KI Reports
Janet Retseck, Alexis Nasr, Yan Lin, Huang Lin, Prateek Mendiratta, Lisa H Butterfield, Ahmad A Tarhini
BACKGROUND: We previously reported early on-treatment significant modulation in circulating regulatory T cell (Treg), myeloid derived suppressor cells (MDSC) and antigen-specific type I CD4+ and CD8+ T cells that correlated with clinical outcome in regionally advanced melanoma patients treated with neoadjuvant ipilimumab. Here, we investigated the long term immunologic impact of CTLA4 blockade. METHODS: Patients were treated with ipilimumab given at 10 mg/kg IV every 3 weeks for 2 doses bracketing surgery...
July 4, 2018: Journal of Translational Medicine
Kathryn D Tuttle, Laurent Gapin
Natural killer T (NKT) cells are a subset of αβ T cells that recognize lipid antigens presented by the nonclassical MHC molecule CD1d. Although numerically small, these cells have been shown to play an important role in the regulation of multiple immune responses, including microbial infection, autoimmunity, and cancer. Even in the steady state, cytokine production by NKT cells influences the basal status and function of other immune cells, including dendritic cells and CD8 T cells. To fully understand their biology and harness them in the clinic, it is imperative to dissect the molecular mechanisms involved in the acquisition of their functionality...
2018: Methods in Molecular Biology
Jiaxiong Tan, Shaohua Chen, Jingying Huang, Youchun Chen, Lijian Yang, Chunli Wang, Jun Zhong, Yuhong Lu, Liang Wang, Kanger Zhu, Yangqiu Li
AIM: The immunosuppressive microenvironment plays a crucial role in T-cell immunodeficiency in multiple myeloma (MM). Overexpression of T-cell immunosuppressive receptors, including programmed death-1 (PD-1) and T-cell immunoglobulin and mucin-domain-containing-3 (Tim-3), may be related to tumor immunosuppression and poor prognosis, and the malignant bone marrow (BM) microenvironment may contribute to such immunosuppression. The purpose of this study was to analyze the distribution of PD-1+ and/or Tim-3+ T cells in different T-cell subset in patients with MM...
June 25, 2018: Asia-Pacific Journal of Clinical Oncology
Elia D Tait Wojno, Celine A Beamer
Innate lymphoid cells (ILCs) comprise a family of innate immune cells that orchestrate mucosal immune responses: initiating, sustaining, and even curbing immune responses. ILCs are relatively rare (≤1% of lymphocytes in mucosal tissues), lack classical cell-surface markers, and can be divided into three subsets (type 1-3 ILCs) based on differences in cytokine production, phenotype, and developmental pathway. Because ILCs can only be identified by combinations of cell-surface markers and cytokine production, multicolor flow cytometry is the most reliable method to purify, characterize, and assess the functionality of ILCs...
2018: Methods in Molecular Biology
Dharmendra Singh, Manisha Ghate, Sheela Godbole, Smita Kulkarni, Madhuri Thakar
Background: CD1d restricted invariant natural killer T (iNKT) cells are important in the activation and regulation of immune responses. Limited information is available regarding the functional role of iNKT cells in the human immunodeficiency virus (HIV) disease progression. Methodology: α-GalCer stimulated iNKT cells were characterized for their functionality in terms of cytokine production (IFN-γ, TNF-α, IL-2, IL-4, and IL-21) and CD107a expression in HIV-1 infected [23 long-term non progressors (LTNPs), 28 progressors, 18 patients before and after suppressive anti-retroviral treatment (ART)] along with 25 HIV-1 negative subjects using multicolor flow cytometry...
2018: Frontiers in Immunology
Jennifer A Juno, Jillian L M Waruk, Kathleen M Wragg, Christine Mesa, Carmen Lopez, Joe Bueti, Stephen J Kent, T Blake Ball, Sandra A Kiazyk
Background: End-stage renal disease (ESRD) is associated with an increased susceptibility to infectious diseases, including infection with Mycobacterium tuberculosis (Mtb) . Mucosal-associated invariant T (MAIT) cells recognize vitamin B metabolites produced by many bacterial species, including Mtb, and may play an important role in providing protective immunity against tuberculosis infection in the lung. To date, little is known about MAIT cell frequency, phenotype, or function in ESRD patients...
2018: Frontiers in Immunology
Xin Wang, Kui-Di Liang, Jun-Ai Zhang, Gan-Bin Liu, Zhi Chen, Chen Chen, Ze-Gang Zhuang, Yu-Qing Liu, Hou-Long Luo, Rui Xi Li, Bi-Ying Zheng, Jun-Fa Xu
B cell activating factor (BAFF), a member of the tumor necrosis factor family, is a key cytokine for B cell survival, a function that is essential for B cell maturation and memory. The expression levels of BAFF and its potential contribution to B cell maturation remain elusive in patients with tuberculous pleural effusion (TPE). The present study enrolled 40 healthy controls (HC) and 45 TPE patients, and investigated the levels of BAFF in the plasma and pleural effusion. Concomitantly, B cell subsets including naïve B cell (CD19+IgD+CD27‑), unswitched B cell (CD19+IgD+CD27+), switched B cell (CD19+IgD‑CD27+), total memory B cell (CD19+CD27+), plasma B cell (CD19+IgD‑CD38+CD27+) and transitional B cell (CD19+IgDdim CD38+) in peripheral blood mononuclear cells (PBMCs) and pleural fluid mononuclear cells (PFMCs) were assessed using multicolor flow cytometry...
August 2018: Molecular Medicine Reports
Joanna Budna, Mariusz Kaczmarek, Agata Kolecka-Bednarczyk, Łukasz Spychalski, Piotr Zawierucha, Joanna Goździk-Spychalska, Michał Nowicki, Halina Batura-Gabryel, Jan Sikora
Cancer metastatic spread to serous cavity causes malignant pleural effusions (MPEs), indicating dismal prognosis. Tumor microenvironment can implement suppressive activity on host immune responses. Thus, we investigated the prevalence of Tregs and the relationship between them and TGF- β and IL-10 concentrations and measured expression of FOXP3 , CTLA-4 , CD28 , and GITR genes, as well as protein expression of selected genes in benign effusions and MPEs. The percentage of Tregs was determined by means of multicolor flow cytometry system...
2018: Journal of Immunology Research
Johannes M Waldschmidt, Praveen Anand, Birgit Knoechel, Jens G Lohr
The presence or absence of minimal residual disease (MRD) in patients with multiple myeloma (MM) has emerged as a useful marker to determine the depth of remission. MRD negativity as an endpoint has been shown to be associated with improved progression-free survival in many studies. MRD detection is therefore part of numerous clinical trial protocols for MM. At the present time, two methodologies are most widely accepted for MRD detection: (1) multicolor flow cytometry and (2) next-generation sequencing-based clonotype detection...
January 2018: Seminars in Hematology
Tali Farine, Michael Parsons, Stephen Lye, Oksana Shynlova
The decidua, also known as the pregnant endometrium, is a critically important reproductive tissue. Decidual cells, comprised mainly of decidualized stromal cells and immune cells, are responsible for the secretion of hormonal and inflammatory factors which are critical for successful blastocyst implantation, placental development and play a role in the initiation of labor at term and preterm. Many pregnancy complications can arise from perturbations of a fine balance of different cell populations comprising decidua...
April 30, 2018: Journal of Visualized Experiments: JoVE
Pascal Ickrath, Norbert Kleinsasser, Xin Ding, Christian Ginzkey, Niklas Beyersdorf, Rudolf Hagen, Thomas Kerkau, Stephan Hackenberg
In patients with chronic rhinosinusitis with nasal polyps (CRSwNP), a relative accumulation of cluster of differentiation (CD)8+ T cells over CD4+ T cells occurs in nasal polyps compared with the peripheral blood. Nasal CD8+ T cells and CD4+ T cells predominantly present an effector memory phenotype. Immunological studies have reported that memory T cells recirculate from the tissues to the peripheral blood and a high percentage of these T cells persist within the tissue. The aim of the present study was to characterize CD69+ sphingosine‑1‑phosphate receptor 1 (S1PR1)‑ tissue resident memory T cells (Trm) in the polyps of patients with CRSwNP...
August 2018: International Journal of Molecular Medicine
Neil N Patel, Michael A Kohanski, Ivy W Maina, Vasiliki Triantafillou, Alan D Workman, Charles C L Tong, Edward C Kuan, John V Bosso, Nithin D Adappa, James N Palmer, De'Broski R Herbert, Noam A Cohen
BACKGROUND: Chronic rhinosinusitis with nasal polyps (CRSwNP) is commonly characterized by type-2 inflammation. It is established that group-2 innate lymphoid cells (ILC2s) are a subset of immune cells important in orchestrating mucosal type-2 response. IL-25 is an epithelial-derived cytokine that is a critical activator of ILC2s. Recent evidence demonstrates that specialized taster epithelial cells, such as solitary chemosensory cells (SCCs), may be producers of IL-25. To elucidate the relationship between SCCs and ILC2s in CRSwNP, we sought to quantify ILC2s and SCCs to determine if these cell types are enriched in nasal polyps compared to healthy sinonasal mucosa...
May 9, 2018: International Forum of Allergy & Rhinology
Hassan Doulabi, Maryam Rastin, Hossein Shabahangh, Ghodratollah Maddah, Abbas Abdollahi, Reza Nosratabadi, Seyed-Alireza Esmaeili, Mahmoud Mahmoudi
T-helper (Th) 22 cells are involved in the immunopathogenesis of inflammatory diseases, but their specific role in the immunopathogenesis of cancer is unknown. In this study, we examined the profile of circulating and intratumoral Th17, Th22 and CD4+ cells co-producing IL-17/IL-22 in colon cancer (CC) patients in relation to tumor staging. Thirty newly diagnosed colon cancer (CC) patients participated in this study. The percentage of Th1 (CD4+ IFN-γ+ IL-17- IL-22- ), Th17 (CD4+ IFN-γ- IL-17+ IL-22- ), Th22 (CD4+ IFN-γ- IL-17- IL-22+ ) and CD4+ cells co-producing IL-17/IL-22 (CD4+ IFN-γ- IL-17+ IL-22+ ) in the peripheral blood, tumor and paratumor tissues was assessed by multicolor flow cytometry...
July 2018: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
Maya Botzman, Irit Gat-Viks
Major efforts are invested in the analysis of phenotypic variation in a population of individuals. While many of these studies focus on the genetic basis of phenotypic variation via measurements of DNA polymorphic sites, the environmental effects are still elusive. Here we propose a methodology, called CCCE ('Cell Composition Common Environment'), to identify environmental effects on the composition of immune cell functionalities. Specifically, CCCE is focused on the common experiences that are shared between siblings (the 'common environment'), designed to correct for cell subpopulation heterogeneity, and is based on a multicolor flow cytometry analysis across a large cohort of human monozygotic and dizygotic twins...
2018: PloS One
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