In vitro Study of Fungus-Like Organisms Peronospora ficariae and Wilsoniana bliti as Potential Inhalant Allergens
Monika Sztandera-Tymoczek1, Sylwia Wdowiak-Wróbel2, Urszula Świderska3
1Department of Virology and Immunology, Institute of Biological Sciences, Maria Curie-Skłodowska University, Lublin, Poland.
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 (Peronospora ficariae) and Albuginales (Wilsoniana bliti) 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 W. bliti extracts did not trigger notable inflammatory responses, P. ficariae 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 P. ficariae 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.
Insights
Certain fungi, like *Peronospora ficariae*, can trigger respiratory allergies by increasing inflammation and damaging airway cells. This study investigated fungal impacts on respiratory health, revealing *P. ficariae* as a potential allergen.
Area of Science:
- Allergy and Immunology
- Microbiology
- Cell Biology
Background:
- Allergic conditions affect nearly 30% of the global population, with fungi being significant respiratory allergens.
- Respiratory allergies are a growing health concern, yet the specific fungal triggers remain largely unidentified.
- Understanding fungal allergens is crucial for developing targeted allergy treatments.
Purpose of the Study:
- To investigate the potential of *Peronospora ficariae* and *Wilsoniana bliti* to induce inflammatory responses in respiratory tract models.
- To explore the role of these fungus-like species in allergic conditions affecting the upper and lower respiratory tracts.
- To elucidate the cellular mechanisms underlying fungal-induced respiratory inflammation.
Main Methods:
- Utilized BEAS-2B and A549 cell lines to model upper and lower respiratory epithelial cells.
- Assessed cytotoxicity using MTT assays and flow cytometry.
- Quantified reactive oxygen species (ROS), proinflammatory cytokines (IL-1β, GM-CSF), and metalloproteinases via flow cytometry and ELISA.
- Evaluated epithelial cell integrity using immunofluorescence.
Main Results:
- *P. ficariae* extracts significantly increased proinflammatory cytokines IL-1β and GM-CSF in both cell lines.
- Increased ROS production and cytokine levels were correlated with the disruption of tight junction proteins (E-cadherin, occludin).
- *W. bliti* extracts did not induce significant inflammatory responses.
Conclusions:
- *P. ficariae* has the potential to compromise the respiratory epithelial barrier.
- The mechanism involves inducing proinflammatory cytokines, ROS, and metalloproteinases, leading to disrupted cell junctions.
- These findings highlight *P. ficariae* as a potential contributor to respiratory allergies.
