Hi all! I'm looking to decrease bacterial uptake by PMNs and I've been using Cytochalasin D with limited success. The bacteria are opsonized, so as far as I know this is driving a CR3-mediated "sinking" phagocytosis. Cytochalasin D is decreasing uptake by less than 50 % on average- 45 min pre-tx and 20 uM being used. So, I'm thinking either my treatment isn't optimal or actin polymerization is less important in complement mediated phagocytosis than in Fc-mediated. What would you use in place of cyto D? Maybe a CD11b block? Suggestions are welcome! TIA.
you can use the blockers like antibodies or you can also use the chemical ligands like heparin sodium salt.
about the Cytochalasin D you need to examine your cells by fluorescent microscope to see weather the used concentration is effective in breaking down of actin and check the cyto toxicity of the Cytochalasin D because it my lead to lysis of your cells if you are not using the best concentration (optimization),
about the intake rate of the bacteria via CR3, have you checked other complement receptors CR1, CR2 and CR4 because there is a cooperation between these receptors during bacterial invasion into the cells, I was using 1ug/ml of Cytochalasin D for 45 m
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