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Measuring Granulocyte and Monocyte Phagocytosis and Oxidative Burst Activity in Human Blood
Published on: September 12, 2016
Simultaneous flow cytometric evaluation of phagocytosis and oxidative burst in human polymorphonuclear cells
M Horvathova1, L Wsolova, E Jahnova
1Department of Clinical Immunology, Research Base of the Slovak Medical University, Bratislava, Slovakia. mira.horvathova@szu.sk
Abstract:
Phagocytosis and oxidative burst (OXIBURST) activity of human polymorphonuclear cells (PMNs) has been simultaneously measured directly in whole blood samples. The ingestion of yeast was assessed by the phagocytosis activity (FA) and phagocytosis index (FI), and the respiratory burst of PMNs was determined as dihydroethidine (DHE) oxidation. We received comparable results in the ingestion of yeast cells by PMNs using either light microscopy (77.31+/-7.56) or flow cytometry detection method (78.26+/-5.14). The significant differences (p<0.05) in FI and OXIBURST activity were find in the patients (2.29+/-0.29 and 14.67+/-3.99, respectively) when compared to healthy donors (1.64+/-0.21 and 32.38+/-14.94, respectively). The two-color flow cytometric procedure permits measurement of two different functions of neutrophils in one step. This flow cytometric procedure is simple, rapid and has the potential to be an alternative assay to test leukocyte function. (Fig. 3, Ref: 30.)
Insights
Human polymorphonuclear cells (PMNs) phagocytosis and oxidative burst (OXIBURST) activity were measured simultaneously in whole blood. Patients showed significantly lower phagocytosis index and OXIBURST activity compared to healthy donors.
Area of Science:
- Immunology
- Cellular Biology
Background:
- Polymorphonuclear cells (PMNs) are crucial for innate immunity.
- Assessing phagocytosis and oxidative burst (OXIBURST) activity is vital for evaluating neutrophil function.
Purpose of the Study:
- To simultaneously measure phagocytosis and OXIBURST activity of human PMNs in whole blood.
- To evaluate a novel two-color flow cytometric assay for leukocyte function testing.
Main Methods:
- Simultaneous measurement of phagocytosis (yeast ingestion) and OXIBURST (dihydroethidine oxidation) in whole blood.
- Utilized flow cytometry and compared results with light microscopy for phagocytosis assessment.
- Assessed phagocytosis activity (FA) and phagocytosis index (FI).
Main Results:
- Flow cytometry provided comparable results to light microscopy for yeast cell ingestion by PMNs.
- Patients exhibited significantly lower phagocytosis index (2.29+/-0.29) and OXIBURST activity (14.67+/-3.99) compared to healthy donors (1.64+/-0.21 and 32.38+/-14.94, respectively).
- The two-color flow cytometric method efficiently measured dual neutrophil functions in a single step.
Conclusions:
- The developed flow cytometric procedure is a simple, rapid, and effective method for assessing PMN phagocytosis and OXIBURST activity.
- This assay has potential as an alternative method for testing leukocyte function.
- Findings indicate impaired neutrophil function in the patient group studied.
