Caveolar transport through nasal epithelium of birch pollen allergen Bet v 1 in allergic patients.
BACKGROUND
Previous work in type I pollen allergies has focused on aberrant immunoresponses.


OBJECTIVE
Our systems-level analyses explore the role of epithelium in early pathogenesis of type I allergic reactions.


METHODS
We began top-down analyses of differences in human nasal epithelial cells and biopsy specimens obtained from patients with birch allergy and healthy control subjects in the resting state and after intranasal in vivo birch pollen challenges. Immunohistochemistry, immunotransmission electron microscopy, mass spectrometry, transcriptomics, and integration of data to a pathway were conducted.


RESULTS
Bet v 1 allergen bound to epithelium immediately after in vivo birch pollen challenge during winter only in allergic individuals. It also travelled through epithelium with caveolae to mast cells. Sixteen unique proteins were found to bind to the Bet v 1 column only in lysates from allergic epithelial cells; 6 of these were caveolar and 6 were cytoskeletal proteins. The nasal epithelial transcriptome analysis from allergic and healthy subjects differed during the winter season, and these subjects also responded differentially to birch pollen challenge. Within this pollen-induced response, the gene ontology categories of cytoskeleton and actin cytoskeleton were decreased in allergic patients, whereas the actin-binding category was enriched in healthy subjects. Integration of microscopic, mass spectrometric, and transcriptomic data to a common protein-protein binding network showed how these were connected to each other.


CONCLUSIONS
We propose a hypothesis of caveolae-dependent uptake and transport of birch pollen allergen in the epithelium of allergic patients only. Application of discovery-driven methodologies can provide new hypotheses worth further analysis of complex multifactorial diseases, such as type I allergy.
Transplantation Laboratory and Infection Biology Research Program, Haartman Institute, University of Helsinki, Helsinki FI-00014, Finland.
Sakari
S
Joenväärä
Pirkko
P
Mattila
Jutta
J
Renkonen
Antti
A
Mäkitie
Sanna
S
Toppila-Salmi
Mikko
M
Lehtonen
Paula
P
Salmi
Satu
S
Lehti
Jarno
J
Mäkinen
Raija
R
Sormunen
Timo
T
Paavonen
Risto
R
Renkonen
D000328:Adult; D052179:Antigens, Plant / Q000235:genetics / Q000276:immunology* / Q000502:physiology; D021941:Caveolae / Q000473:pathology / Q000502:physiology*; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D013058:Mass Spectrometry; D009297:Nasal Mucosa / Q000473:pathology / Q000502:physiology*; D011994:Recombinant Proteins / Q000235:genetics; D012015:Reference Standards; D006255:Rhinitis, Allergic, Seasonal; D055815:Young Adult
D052179:Antigens, Plant; D011994:Recombinant Proteins