Non‐typeable Haemophilus influenzae (NTHi) invades host cells by binding of the platelet‐activating factor (PAF) receptor via lipooligosaccharide (LOS) glycoforms containing phosphorylcholine (ChoP). ...The effect of NTHi infection on host cell signalling and its role in NTHi invasion was examined. The infection of human bronchial epithelial cells with NTHi 2019 increased cytosolic Ca2+ levels, and the invasion of bronchial cells by NTHi 2019 was inhibited by pretreatment with the cell‐permeant intracellular Ca2+ chelator BAPTA‐AM (P = 0.022) or thapsigargin (P = 0.016). Cytosolic inositol phosphate (IP) levels were also increased after infection with NTHi 2019 (P < 0.001), but not after infection with isogenic mutants expressing altered LOS glycoforms lacking ChoP. PAF receptor antagonist reduced NTHi 2019‐stimulated IP production in a dose‐dependent manner. NTHi 2019 invasion was inhibited by pertussis toxin (PTX) and the phosphatidylinositol‐3‐kinase inhibitors wortmannin and LY294002. The less invasive strain NTHi 7502 also initiated IP production, but was unaffected by PAF receptor antagonist or PTX. These data demonstrate that the binding of the PAF receptor by NTHi initiates receptor coupling to a PTX‐sensitive heterotrimeric G protein complex, resulting in a multifactorial host cell signal cascade and bacterial invasion. Moreover, the data suggest that NTHi strains initiate cell signalling and invade by different mechanisms, and that invasion mediated by PAF receptor activation is more efficient than macropinocytosis.
Neisseria meningitidisis an organism whose environmental niche is limited to the human host. It can frequently colonize the human nasopharynx and has the ability to cause severe systemic infections. ...These infections can be sporadic, endemic or occur in outbreaks associated with more virulent meningococcal strains. Studies have demonstrated that the meningococcus can form biofilms both in vivo and ex vivo. In this chapter, we discuss methods to establish biofilms in the laboratory for in-depth biochemical, genetic, or microscopic studies.
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