Mycoplasma pneumoniae is an obligate pathogen that causes pneumonia, tracheobronchitis, pharyngitis and asthma in humans. It is well recognized that membrane lipoproteins are immunostimulants exerting as lipopolysaccharides (LPS) and play a crucial role in the pathogenesis of inflammatory responses upon M. pneumoniae infection. Here, we report that the M. pneumoniae-derived lipids are another proinflammatory agents. Using an antibody-neutralizing assay, RNA interference or specific inhibitors, we found that Toll-like receptor 4 (TLR-4) is essential for M. pneumoniae lipid-induced tumour necrosis factor (TNF)-α and interleukin (IL)-1β production. We also demonstrate that NLR family pyrin domain containing 3 inflammasome (NLRP3) inflammasome, autophagy and nuclear factor kappa B (NF-κB)-dependent pathways are critical for the secretion of proinflammatory cytokines, while inhibition of TLR-4 significantly abrogates these events. Further characterization revealed that autophagy-mediated inflammatory responses involved the activation of NF-κB. In addition, the activation of NF-κB promoted lipid-induced autophagosome formation, as revealed by assays using pharmacological inhibitors, 3-methyladenine (3-MA) and Bay 11-7082, or silencing of atg5 and beclin-1. These findings suggest that, unlike the response to lipoprotein stimulation, the inflammation in response to M. pneumoniae lipids is mediated by the TLR-4 pathway, which subsequently initiates the activation of NLRP3 inflammasome and formation of a positive feedback loop between autophagy and NF-κB signalling cascade, ultimately promoting TNF-α and Il-1β production in macrophages.
Keywords: Mycoplasma pneumonia; NF-κB; NLRP3; Toll-like receptor 4; autophagy; lipid.
© 2020 British Society for Immunology.