Camel Granulocyte ROS Production Assay

The production of ROS metabolites was measured in 96-well round-bottom microtiter plates (Corning, NY, USA) as previously described [29 (link)]. Camel granulocytes (1 × 10⁶/100 μL/well) were incubated in 50 μL RPMI culture medium alone or in a medium containing T. evansi (4 × 10⁶ parasite/mL) for 30 min (37 °C, 5% CO₂). After 15 min of incubation, dihydrorhodamine (DHR) 123 (Mobitec, Goettingen, Germany) was added to the cells at a final concentration of 750 ng/mL. The cells were washed in RPMI medium, and ROS production was analyzed by flow cytometry.

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Hussen J., AL-Jabr O.A., Alkuwayti M.A., Alrabiah N.A., Falemban B., Alouffi A., Al Salim W.S., Kamyingkird K, & Desquesnes M. (2023). A Flow Cytometry Study of the Binding and Stimulation Potential of Inactivated Trypanosoma evansi toward Dromedary Camel Leukocytes. Pathogens, 13(1), 21.

Publication 2023

96-well round-bottom microtiter plates DHR) 123

Corresponding Organization : King Faisal University

Other organizations : King Abdulaziz City for Science and Technology, Kasetsart University, Biologie et Génétique des Interactions Plante-Parasite

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Variable analysis

independent variables

Presence of T. evansi (4 × 10^6 parasites/mL)

dependent variables

Production of ROS metabolites

control variables

Camel granulocytes (1 × 10^6/100 μL/well)
RPMI culture medium
Incubation time (30 min)
Incubation temperature (37 °C)
Incubation atmosphere (5% CO2)
Concentration of dihydrorhodamine (DHR) 123 (750 ng/mL)

controls

Camel granulocytes incubated in RPMI culture medium alone (negative control)

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