In vitro Study of Fungus-Like Organisms and as Potential Inhalant Allergens.

PURPOSE

Allergic conditions have surged to epidemic proportions globally, affecting almost 30% of people worldwide. Fungi are a significant source of allergens, contributing to up to 6% of respiratory illnesses in the general population. Nonetheless, the exact cause of respiratory allergies often remains elusive. This research sought to explore the potential of two fungus-like species from the Peronosporales () and Albuginales () to induce a proinflammatory response in vitro models of the upper and lower respiratory tract.

MATERIALS AND METHODS

BEAS-2B and A549 cell lines were used to mimic upper and lower respiratory epithelial cells. The cytotoxicity of fungal extracts was assessed using MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) and flow cytometry assays. Reactive oxygen species (ROS) production in the cells was measured through flow cytometry, while ELISA (enzyme-linked immunosorbent assay) tests were used to quantify the production of proinflammatory cytokines and metalloproteinases. Immunofluorescence techniques were utilized to evaluate markers of cell integrity.

RESULTS

Although extracts did not trigger notable inflammatory responses, significantly enhanced the production of proinflammatory cytokines IL-1β and GM-CSF in both cell lines, which are linked to allergic reactions. The rise in these cytokines and increased ROS production were associated with disrupting tight junction proteins, such as E-cadherin and occludin, in epithelial cells.

CONCLUSION

The results indicate that extracts have the potential to collectively compromise the epithelial barrier in the upper and lower respiratory tracts by inducing proinflammatory cytokines and promoting the production of reactive oxygen species and metalloproteinases. While none of these parameters were exceptionally high, their combined effect was observed to disrupt epithelial cell junctions.