Journal: Frontiers in Immunology
Article Title: The Inhibitory Effect of Curosurf® and Alveofact® on the Formation of Neutrophil Extracellular Traps
Figure Lengend Snippet: Decrease of P2Y6 Receptor and Calcium mobilization after treatment without change in intracellular PAD4 levels. (A) Neutrophils were treated as described before. P2Y6 was measured by flow cytometry with a FITC-conjugated antibody. Treatment with Alveofact ® decreases extracellular expression of the P2Y6 receptor independent of timing or dosing, while Curosurf ® does not exhibit this effect in every dosing. (B) Calcium levels at 3 h after treatment were used. Data were normalized to stimulated PMA control due to the heterogeneity in basic calcium levels and the response of the neutrophils to stimulation. Results are similar to the findings of the P2Y6 receptor expression. (C) Intracellular PAD4 levels were determined by ELISA using cell lysates of 1×10 6 cells. Interestingly, PAD4 levels did not change after treatment with surfactant compared to PMA-stimulated neutrophils. Significance level was set as p
Article Snippet: Cells were washed twice with PBS (Thermo Fischer, Waltham, MA, USA) and labeled with propidium iodide (PI) and Annexin-V-FITC (Becton, Dickinson and Company, Franklin Lakes, NJ, USA) or anti-CD11b-VioBlue (mAb REA713, IgG1) and anti-CD66b-APC (mAb REA306, IgG1) antibodies (Miltenyi Biotec, Bergisch Gladbach, Germany) or anti-P2Y6 Receptor-FITC (Alomone Labs, Jerusalem, Israel) as described in the manufacturer’s protocols.
Techniques: Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay