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Macrophages mediated antibacterial responses are impaired by selectively inhibiting the activity of p300/CREB-binding protein

Li Yiping  Li Gang  Fang Fang  Zhou Huiting  Li Zhiheng  Pan Jian  Li Yanhong  Feng Xing  
【摘要】:Objective:C646 is a newly discovered competitive p300/CREB-binding protein-specific inhibitor.Studies have shown its potential antitumor activity,but the effect on host defenses remains largely unknown.Methods:Here we investigated the immunomodulatory and anti-bacterial activity of C646 on mouse macrophages in vitro.The effect of C646 on the expression of inflammatory cytokines was detected by flow cytometry and real-time PCR.The effect of C646 on intracellular signaling pathway was detected by western blot.The effect of C646 on the binding of LPS to macrophages,phagocytic receptor expression,and the ability of bacterial uptake and phagocytosis was monitored by flow cytometry.The effect of C646 on phagosomal maturation was observed by confocal fluorescence microscopy.The effect of C646 on macrophage bactericidal ability was detected by in vitro assay.Results:(1) The expression levels of TNF-a,MCP-1 and IL-6 in macrophages were inhibited by C646.The levels of CD14 and TLR4/MD2 were inhibited,but no obvious effect was found in the binding ability of LPS to macrophages.C646 also inhibits JNK,ERK1/2,and p65 phosphorylation,but not p38 phosphorylation.(2) The expression levels of FcγRⅢ/Ⅱ and CR3 on macrophages were decreased by C646,but no significant effect on the level of FcγRI.C646 can also significantly inhibit macrophage uptake and phagocytosis of E.coli,and block its phagosome-lysosome fusion.Moreover,macrophage bactericidal activity was reduced by C646.Conclusion:Macrophage-mediated inflammatory response and the control of bacterial pathogens could be impaired by C646.Further study is needed before C646 used in cancer therapy.

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