Alison Scaife

Maternal intake of antioxidant vitamins in pregnancy in relation to maternal and fetal plasma levels at delivery

The aim of the present study was to test the hypothesis that maternal intake of antioxidant vitamins is associated with maternal and cord plasma levels at delivery. Women were recruited in early pregnancy in Aberdeen Maternity Hospital and habitual diet during pregnancy was assessed by a food-frequency questionnaire mailed at 34 weeks gestation. Blood samples were taken at recruitment (n 1149) and maternal (n 1149) and cord blood samples (n 747) taken at delivery for analyses of vitamins A, C, E and beta-carotene. Maternal plasma levels of vitamin E and beta-carotene at delivery were significantly higher than levels in early pregnancy while levels of vitamins A and C were significantly lower. Positive correlations were observed for maternal levels of all the vitamins between early pregnancy and delivery. At delivery, maternal plasma concentrations of vitamins A, E and beta-carotene were significantly higher than cord levels, while maternal levels of vitamin C were significantly lower. There were significant correlations between maternal and cord plasma concentrations for beta-carotene and vitamin C but not for vitamins A or E. Maternal dietary intakes were positively correlated with maternal plasma levels of vitamins C, E and beta-carotene in early pregnancy, with maternal plasma levels of beta-carotene and vitamin C at delivery and with cord plasma levels of beta-carotene and vitamin C. The results from the present study show that, in this population, maternal diet influences cord plasma levels of beta-carotene and vitamin C, but not vitamins A and E.

Maternal vitamin D and E intakes during early pregnancy are associated with airway epithelial cell responses in neonates

Antenatal factors including maternal diet may predispose to airway disease, possibly by impacting on fetal airway development.

Primary Paediatric Bronchial Airway Epithelial Cell in Vitro Responses to Environmental Exposures

The bronchial airway epithelial cell (BAEC) is the site for initial encounters between inhaled environmental factors and the lower respiratory system. Our hypothesis was that release of pro inflammatory interleukins (IL)-6 and IL-8 from primary BAEC cultured from children will be increased after in vitro exposure to common environmental factors. Primary BAEC were obtained from children undergoing clinically indicated routine general anaesthetic procedures. Cells were exposed to three different concentrations of lipopolysaccharide (LPS) or house dust mite allergen (HDM) or particulates extracted from side stream cigarette smoke (SSCS). BAEC were obtained from 24 children (mean age 7.0 years) and exposed to stimuli. Compared with the negative control, there was an increase in IL-6 and IL-8 release after exposure to HDM (p ≤ 0.001 for both comparisons). There was reduced IL-6 after higher compared to lower SSCS exposure (p = 0.023). There was no change in BAEC release of IL-6 or IL-8 after LPS exposure. BAEC from children are able to recognise and respond in vitro with enhanced pro inflammatory mediator secretion to some inhaled exposures.

Inhibitory effects of Montelukast on mediator release by nasal epithelial cells from asthmatic subjects with or without allergic rhinitis

AIMS This study tested inhibitory effects of in vitro Montelukast treatment on nasal airway epithelial cells (AEC) cultured from asthmatic patients treated with Montelukast with and without concomitant allergic rhinitis. We further examined the effect of Montelukast withdrawal in these patients on cytokine release from cultured nasal AEC. METHODS Nasal AEC were collected by brushings from subjects with a history of stable (no exacerbations or change in medication for ≥ 1 month) physician confirmed mild/moderate asthma whose asthma symptoms were documented to benefit from Montelukast treatment (NCT01230437). Release of the following mediators by nasal AEC were measured: IL-8, IL-6, IL-10, GM-CSF, RANTES, eotaxin and IFN-γ. Nasal AEC were cultured before and one week after withdrawal of their Montelukast treatment. RESULTS Forty two asthmatics were recruited. Nasal AEC were successfully cultured in 17 at the first assessment, 14 at the second assessment and in 10 individuals at both assessments. Nasal AEC release was no different between asthmatics with or without allergic rhinitis. Montelukast significantly suppressed the release of IL-8 (p = 0.016), IL-6 (p = 0.006), RANTES (p = 0.002) and IFN-γ (p = 0.046), in a dose dependent manner in unstimulated cultures but not in those stimulated with IL-1/TNF. Withdrawal of Montelukast treatment, was associated with increased IL-8 and RANTES secretion in unstimulated nasal AEC cultured from subjects with asthma and allergic rhinitis but not with asthma alone. CONCLUSIONS Montelukast treatment for asthma symptoms reversibly suppresses nasal AEC release of pro-inflammatory mediators (i.e. IL-8 and RANTES) but only in those cells cultured from subjects with concomitant allergic rhinitis.

Culture of Airway Epithelial Cells from Neonates Sampled within 48-Hours of Birth

Introduction Little is known about how neonatal airway epithelial cell phenotype impacts on respiratory disease in later life. This study aimed to establish a methodology to culture and characterise neonatal nasal epithelial cells sampled from healthy, non-sedated infants within 48 hours of delivery. Methods Nasal epithelial cells were sampled by brushing both nostrils with an interdental brush, grown to confluence and sub-cultured. Cultured cells were characterised morphologically by light and electron microscopy and by immunocytochemistry. As an exemplar pro-inflammatory chemokine, IL-8 concentrations were measured in supernatants from unstimulated monolayers and after exposure to IL-1β/TNF-α or house dust mite extract. Results Primary cultures were successfully established in 135 (91%) of 149 neonatal samples seeded, with 79% (n  =  117) successfully cultured to passage 3. The epithelial lineage of the cells was confirmed by morphological analysis and immunostaining. Constitutive IL-8 secretion was observed and was upregulated by IL-1β/TNF-α or house dust mite extract in a dose dependent manner. Conclusion We describe a safe, minimally invasive method of culturing nasal epithelial cells from neonates suitable for functional cell analysis offering an opportunity to study “naïve” cells that may prove useful in elucidating the role of the epithelium in the early origins of asthma and/or allergic rhinitis.

Pro-inflammatory mediator responses from neonatal airway epithelial cells and early childhood wheeze

Airway epithelial cell (AEC) function differs between children with and without asthma. Here, we associated neonatal AEC function with asthma symptoms at 4 years of age.

Novel in-vitro model to study first responses of airway epithelial cells to allergen and pro-inflammatory stimuli at birth

Abstract Background The airway epithelium is increasingly being implicated in the pathogenesis of asthma. Although believed to be important, little is known about how the neonatal airway epithelial cell (AEC) phenotype impacts on respiratory disease in later life. The aim of this study was to establish a methodology for culturing neonatal nasal AEC and to describe AEC response in vitro. Methods AECs were sampled from healthy, unsedated infants during the first week of life by brushing both nostrils with an interdental brush. Sampled AECs were used for cytospin preparation or grown to confluence before subculture. Cultured cells were characterised morphologically and by immunocytochemistry. Interleukin-8 concentrations were measured in supernatants from monolayers at rest and after exposure to concentration ranges of interleukin 1β and tumour necrosis factor α or house dust mite extract. Findings Primary cultures were successfully established in 109 (92%) of 117 neonates sampled, with 93 (80%) successfully cultured to confluence at third passage. The epithelial lineage of the cells was confirmed by morphological analysis and immunocytochemistry. Constitutive interleukin-8 secretion was observed and was upregulated by both stimuli in a dose dependent manner. Interpretation We describe a safe, minimally invasive method of culturing AECs from neonates suitable for functional cell analysis and amenable to large population based studies. This novel technique offers a unique opportunity to study naive AECs not yet exposed to the modifying effects of environmental pollutants and viral pathogens and may prove useful in elucidating the early origins of asthma. Funding Chief Scientist Office of the Scottish Government.