Intracellular Cytokine Analysis of Blood and BAL Samples

Directly after collection, blood and BAL samples were added to an equal volume (100 μL/tube) to IMDM (Iscove’s Modified Dulbecco’s Media, Thermo Fisher Scientific, Waltham, MA, USA) with or without 200 ng/mL of Escherichia coli LPS (E. coli K12 LPS, Invivogen, San Diego, CA, USA), then incubated for 4 h at 37 °C and 5% CO₂ [23 (link)]. Samples drawn to measure the intracellular cytokine synthesis of TNF-α were incubated with a Golgi inhibitor (GolgiStop™ 2 μM, Becton Dickinson, San Jose, CA, USA) for 4 h at 37 °C and 5% CO₂.

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Bendib I., Beldi-Ferchiou A., Schlemmer F., Maitre B., Surenaud M., Hüe S., Carteaux G., Razazi K., Lelièvre J.D., Lévy Y., Mekontso Dessap A., Godot V, & de Prost N. (2021). Functional Ex Vivo Testing of Alveolar Monocytes in Patients with Pneumonia-Related ARDS. Cells, 10(12), 3546.

Publication 2021

Iscove’s Modified Dulbecco’s Media, E. coli K12 LPS,

Blood Cytokine E coli k12 Escherichia coli Golgi Intracellular Synthesis Tnf α

Corresponding Organization : Institut Mondor de Recherche Biomédicale

Other organizations : Institut de Recherche Vaccinale, Hôpital Albert-Chenevier

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Variable analysis

independent variables

Addition of 200 ng/mL of Escherichia coli LPS (E. coli K12 LPS) to the samples

dependent variables

Intracellular cytokine synthesis of TNF-α

control variables

Blood and BAL samples were added to an equal volume (100 μL/tube) of IMDM (Iscove's Modified Dulbecco's Media)
Incubation time of 4 h at 37 °C and 5% CO2

controls

Positive control: Samples incubated with 200 ng/mL of E. coli LPS
Negative control: Samples incubated without E. coli LPS

Annotations

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