Monte Carlo studies of two-dimensional polymer–solvent systems

<div><p>Allergic bronchopulmonary aspergillosis (ABPA) in asthma is a severe, life-affecting disease that potentially affects over 4.8 million people globally. In the UK, ABPA is predominantly caused by the fungus <i>Aspergillus fumigatus</i>. Phagocytosis is important in clearance of this fungus, and Early Endosome Antigen 1 (<i>EEA1</i>) has been demonstrated to be involved in phagocytosis of fungi. We sought to investigate the role of <i>EEA1</i> mutations and phagocytosis in ABPA. We used exome sequencing to identify variants in <i>EEA1</i> associated with ABPA. We then cultured monocyte-derived macrophages (MDMs) from 17 ABPA subjects with <i>A</i>. <i>fumigatus</i> conidia, and analyzed phagocytosis and phagolysosome acidification in relation to the presence of these variants. We found that variants in <i>EEA1</i> were associated with ABPA and with the rate of phagocytosis of <i>A</i>. <i>fumigatus</i> conidia and the acidification of phagolysosomes. MDMs from ABPA subjects carrying the disease associated genotype showed increased acidification and phagocytosis compared to those from ABPA subjects carrying the non-associated genotypes or healthy controls.The identification of ABPA-associated variants in EEA that have functional effects on MDM phagocytosis and phagolysosome acidification of <i>A</i>. <i>fumigatus</i> conidia revolutionizes our understanding of susceptibility to this disease, which may in future benefit patients by earlier identification or improved treatments. We suggest that the increased phagocytosis and acidification observed demonstrates an over-active MDM profile in these patients, resulting in an exaggerated cellular response to the presence of <i>A</i>. <i>fumigatus</i> in the airways.</p></div

<i>EEA1</i> mutations associated with ABPA.

<p><i>EEA1</i> mutations associated with ABPA.</p

<i>EEA1</i> expression, phagocytosis and phagolysosome acidification after siRNA treatment.

<p>Mean and SEM shown. Groups compared by t-test. A) siRNA treatment. B) Phagocytosis. C) Phagolysosome acidification.</p

Mutations in <i>EEA1</i> are associated with allergic bronchopulmonary aspergillosis and affect phagocytosis of <i>Aspergillus fumigatus</i> by human macrophages - Fig 3

<p><b>Association of EEA1 genotype (A) and correlation of disease-associated alleles (B) with the percentage of conidial phagocytosis observed in MDMs after co-culture with <i>A</i>. <i>fumigatus</i>.</b> A) Median and interquartile ranges are shown. For the ABPA groups, each point represents an individual. For the healthy group, which is shown for interest, each point represents a replicate experiment using the same healthy subject. ABPA groups were compared by Mann-Whitney tests. Healthy groups were not compared as they only contained data for one individual, and are shown for interest only. B) Only ABPA subjects are shown, and each point represents an individual. Correlation was calculated using the Pearson R test.</p

Mutations in <i>EEA1</i> are associated with allergic bronchopulmonary aspergillosis and affect phagocytosis of <i>Aspergillus fumigatus</i> by human macrophages - Fig 2

<p><b>Association of EEA1 genotype (A) and correlation of disease-associated alleles (B) with the percentage of phagolysosome acidification observed in MDMs after co-culture with <i>A</i>. <i>fumigatus</i>.</b> A) Median and interquartile ranges are shown. For the ABPA groups, each point represents an individual. For the healthy group, which is shown for interest, each point represents a replicate experiment using the same healthy subject. ABPA groups were compared by Mann-Whitney tests. Healthy groups were not compared as they only contained data for one individual, and are shown for interest only. B) Only ABPA subjects are shown, and each point represents an individual. Correlation was calculated using the Pearson R test.</p