title: Pneumococcal infection of respiratory cells exposed to welding fumes; Role of oxidative stress and HIF-1 alpha.
abstract: Welders are more susceptible to pneumococcal pneumonia. The mechanisms are yet unclear. Pneumococci co-opt the platelet activating factor receptor (PAFR) to infect respiratory epithelial cells. We previously reported that exposure of respiratory cells to welding fumes (WF), upregulates PAFR-dependent pneumococcal infection. The signaling pathway for this response is unknown, however, in intestinal cells, hypoxia-inducible factor-1 α (HIF 1α) is reported to mediate PAFR-dependent infection. We sought to assess whether oxidative stress plays a role in susceptibility to pneumococcal infection via the platelet activating factor receptor. We also sought to evaluate the suitability of nasal epithelial PAFR expression in welders as a biomarker of susceptibility to infection. Finally, we investigated the generalisability of the effect of welding fumes on pneumococcal infection and growth using a variety of different welding fume samples. Nasal epithelial PAFR expression in welders and controls was analysed by flow cytometry. WF were collected using standard methodology. The effect of WF on respiratory cell reactive oxygen species production, HIF-1α expression, and pneumococcal infection was determined using flow cytometry, HIF-1α knockdown and overexpression, and pneumococcal infection assays. We found that nasal PAFR expression is significantly increased in welders compared with controls and that WF significantly increased reactive oxygen species production, HIF-1α and PAFR expression, and pneumococcal infection of respiratory cells. In unstimulated cells, HIF-1α knockdown decreased PAFR expression and HIF-1α overexpression increased PAFR expression. However, in knockdown cells pneumococcal infection was paradoxically increased and in overexpressing cells infection was unaffected. Nasal epithelial PAFR expression may be used as a biomarker of susceptibility to pneumococcal infection in order to target individuals, particularly those at high risk such as welders, for the pneumococcal vaccine. Expression of HIF-1α in unexposed respiratory cells inhibits basal pneumococcal infection via PAFR-independent mechanisms.
authors.affiliation: Centre for Genomics and Child Health, Blizard Institute for Cell and Molecular Sciences, London, United Kingdom.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Grigg
authors.affiliation: Centre for Genomics and Child Health, Blizard Institute for Cell and Molecular Sciences, London, United Kingdom.
authors.forename: Lisa
authors.initials: L
authors.lastname: Miyashita
authors.affiliation: Centre for Genomics and Child Health, Blizard Institute for Cell and Molecular Sciences, London, United Kingdom.
authors.forename: Reetika
authors.identifier: http://orcid.org/0000-0001-5509-2502
authors.initials: R
authors.lastname: Suri
mesh_terms: D000293:Adolescent; D000328:Adult; D016022:Case-Control Studies; D005500:Follow-Up Studies; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D016273:Occupational Exposure / Q000009:adverse effects*; D018384:Oxidative Stress; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism; D011008:Pneumococcal Infections / Q000209:etiology* / Q000378:metabolism / Q000473:pathology; D011379:Prognosis; D017382:Reactive Oxygen Species / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D020545:Respiratory Mucosa / Q000382:microbiology*; D013296:Streptococcus pneumoniae / Q000472:pathogenicity*; D014896:Welding / Q000379:methods*; D055815:Young Adult
chemical_list: C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D010980:Platelet Membrane Glycoproteins; D017382:Reactive Oxygen Species; D043562:Receptors, G-Protein-Coupled; C039351:platelet activating factor receptor