A rapid flow cytometry assay for the assessment of calcium mobilization in human neutrophils in a small volume of lysed whole-blood

2009 ◽  
Vol 340 (2) ◽  
pp. 154-157 ◽  
Author(s):  
Philippe Desmeules ◽  
Maurice Dufour ◽  
Maria J.G. Fernandes
Keyword(s):  
Flow Cytometry ◽  
Whole Blood ◽  
Small Volume ◽  
Calcium Mobilization ◽  
Human Neutrophils ◽  
Flow Cytometry Assay ◽  
Rapid Flow

APPLICATION OF A KINETIC WHOLE BLOOD FLOW CYTOMETRY METHOD TO STUDY FREE CALCIUM MOBILIZATION IN PLATELETS OF HYPERTENSIVE PATIENTS. EFFECT OF DOXAZOSIN

2004 ◽  
Vol 22 (Suppl. 2) ◽  
pp. S126
Author(s):  
M. Labiós Gómez ◽  
M. Martinez Silvestre ◽  
F. Gabriel Botella ◽  
V. Guiral Olivan ◽  
S. Palanca Suela ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Blood Flow ◽  
Whole Blood ◽  
Calcium Mobilization ◽  
Free Calcium ◽  
Hypertensive Patients

scholarly journals Ultra-rapid flow cytometry assay for colistin MIC determination in Enterobacterales, Pseudomonas aeruginosa and Acinetobacter baumannii

2020 ◽  
Vol 26 (11) ◽  
pp. 1559.e1-1559.e4
Author(s):  
Daniela Fonseca e Silva ◽  
Ferdinando F. Andrade ◽  
Rosário Gomes ◽  
Ana Silva-Dias ◽  
Inês Martins-Oliveira ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Pseudomonas Aeruginosa ◽  
Acinetobacter Baumannii ◽  
Flow Cytometry Assay ◽  
Rapid Flow

scholarly journals Use of Flow Cytometry to Evaluate Phagocytosis of Staphylococcus aureus by Human Neutrophils

2021 ◽  
Vol 12 ◽  
Author(s):  
Elena Boero ◽  
Iris Brinkman ◽  
Thessely Juliet ◽  
Eline van Yperen ◽  
Jos A. G. van Strijp ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Flow Cytometry ◽  
Immune Responses ◽  
Human Neutrophils ◽  
Host Proteins ◽  
Flow Cytometry Assay ◽  
Immune Therapies ◽  
Human Immune Response ◽  
Human Immune ◽  
Phagocytic Uptake

Neutrophils play a key role in the human immune response to Staphylococcus aureus infections. These professional phagocytes rapidly migrate to the site of infection to engulf bacteria and destroy them via specialized intracellular killing mechanisms. Here we describe a robust and relatively high-throughput flow cytometry assay to quantify phagocytosis of S. aureus by human neutrophils. We show that effective phagocytic uptake of S. aureus is greatly enhanced by opsonization, i.e. the tagging of microbial surfaces with plasma-derived host proteins like antibodies and complement. Our rapid assay to monitor phagocytosis can be used to study neutrophil deficiencies and bacterial evasion, but also provides a powerful tool to assess the opsonic capacity of antibodies, either in the context of natural immune responses or immune therapies.

Human monocytes express functional receptors for granulocyte colony– stimulating factor that mediate suppression of monokines and interferon-γ

Blood ◽  
2000 ◽  
Vol 95 (1) ◽  
pp. 270-276 ◽  
Author(s):  
Eva-Maria Boneberg ◽  
Lars Hareng ◽  
Florian Gantner ◽  
Albrecht Wendel ◽  
Thomas Hartung
Keyword(s):  
Flow Cytometry ◽  
Whole Blood ◽  
Receptor Expression ◽  
Human Neutrophils ◽  
Enzyme Linked Immunosorbent Assay ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Ifn Γ ◽  
Stimulating Factor

In a double-blind, placebo-controlled, randomized study, 10 healthy men received either a single dose of 480 μg granulocyte colony-stimulating factor (G-CSF) or saline. Blood taken from the volunteers was stimulated with 10 μg/mL endotoxin and released cytokines were measured by enzyme-linked immunosorbent assay. Expression of G-CSF receptors on leukocytes was examined by flow cytometry and reverse transcriptase-polymerase chain reaction. Functional activity of these receptors was tested by challenging isolated leukocyte populations to release cytokines with endotoxin in the presence of G-CSF. The G-CSF treatment attenuated the release of the proinflammatory cytokines tumor necrosis factor (TNF)-, interleukin (IL)-12, IL-1β, and interferon (IFN)-γ in ex vivo lipopolysaccharide (LPS)-stimulated whole blood. In blood from untreated volunteers the presence of G-CSF in vitro also attenuated the LPS-stimulated release of these cytokines. G-CSF in vitro also attenuated TNF- release from elutriation-purified monocytes. In the presence of 10 ng/mL recombinant TNF-, the attenuation of LPS-inducible IFN-γ release by G-CSF was blunted in whole blood. However, G-CSF had no such effect on IFN-γ release from isolated lymphocytes stimulated with anti-CD3 or a combination of TNF- and IL-12. G-CSF receptor expression was detected in human neutrophils and monocytes but not in lymphocytes by means of RT-PCR as well as flow cytometry. These results indicate that G-CSF receptors expressed on monocytes are functional in modulating monokine release. We conclude that the attenuation of IFN-γ release from lymphocytes is not a direct effect of G-CSF on these cells but is rather due to the inhibition of monocytic IL-12 and TNF- release by G-CSF. (Blood. 2000;95:270-276)

Sign in / Sign up

Export Citation Format

Share Document