Nasal Epithelial Cells Activated with Alternaria and House Dust Mite Induce Not Only Th2 but Also Th1 Immune Responses

Abstract

Chronic rhinosinusitis (CRS) is a heterogeneous disease characterized by mucosal inflammation. Airborne allergens are associated with upper and lower airway inflammatory disease. We investigated the effects of airborne allergen stimulation in the nasal epithelial cells and their effect on the peripheral blood mononuclear cells’ (PBMCs) Th immune polarization. Interleukin (IL)-10, IL-25, IL-33, and thymic stromal lymphopoietin (TSLP) levels were determined using the enzyme-linked immunosorbent assay (ELISA) in nasal polyp tissues. Cultured primary nasal epithelial cells were stimulated with Alternaria alternata, Aspergillus fumigatus, Dermatophagoides pteronyssinus (DP), and Dermatophagoides farina (DF) for 48 hours. IL-6, IL-25, IL-33, and TSLP production were measured by ELISA, and the nuclear factor-κB (NF-κB), activator protein 1 (AP-1), and mitogen-activated protein kinase (MAPK) expression were determined by western blot analyses. PBMCs were cultured with nasal epithelial cell-conditioned media (NECM), and IL-5, interferon (IFN)-γ, and tumor necrosis factor (TNF)-α were measured. Innate lymphoid type2 cells (ILC2) were analyzed with flowcytometry. IL-25, IL-33, and TSLP levels were significantly higher in eosinophilic nasal polyps. Alternaria, DP, and DF enhanced IL-33 and TSLP production from the nasal epithelial cells through the NF-κB, AP-1, and MAPK pathway. NECM induced IL-5, IFN-γ, and TNF-α production from PBMCs, without increasing ILC2 expression. Alternaria and house dust mites enhanced the chemical mediator production from nasal epithelial cells and these allergens may induce not only Th2 inflammatory responses but also Th1 inflammatory responses in the nasal mucosa.

A scoping review: What are the cellular mechanisms that drive the allergic inflammatory response to fungal allergens in the lung epithelium?

“…The few studies ( n = 9, 15%) that compared different fungal species suggest that they can induce different responses in airway epithelia. For example, while 48 h exposure to both Aspergillus and Alternaria extracts stimulated IL6 in primary HNE cells, only Alternaria extract enhanced IL33 and TSLP 85 . A similar induction of IL6 and lack of IL33 or TSLP was observed following a 24 h exposure to Aspergillus extract in a separate study using mouse lung epithelial cells 73 .…”

“…IL33 is constitutively expressed in epithelial cells within the lung and resides, under normal conditions, in the cell nucleus in its full length form 11,112 . Expression of IL33 was shown to be mediated by nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF‐κB), activator protein 1 (AP‐1) and MAPK transcription factors 85 . Indeed, IL33 was confirmed to be localised within the nuclei of HBE cells at rest 63 .…”

Asian Sand Dust Particles Enhance the Development of Aspergillus fumigatus Biofilm on Nasal Epithelial Cells

“…Specimens were placed in Ham’s F-12 medium supplemented with 100 IU of penicillin, 100 µg/mL streptomycin, and 2 µg/mL amphotericin B, and were then transported to the laboratory. Primary nasal epithelial cells were isolated using 0.1% dispase (Roche Diagnostics, Mannheim, Germany), as previously described [ 25 ]. Cell suspensions (1 × 10 6 cells/mL) were plated in 6-well culture plates (140675, Thermo Fisher Scientific, Waltham, MA, USA) and grown to 80–90% confluence on culture plates in a 5% CO 2 humidified incubator at 37 °C.…”

Interactions of thymic stromal lymphopoietin with interleukin-4 in adaptive immunity during Aspergillus fumigatus keratitis

“…The innate immune system identifies pathogen-associated molecular patterns (PAMPs) through pattern recognition receptors [6][7][8][9][10] . Aspergillus fumigatus (AF) can induce the production of inflammatory factors through nuclear factor κB (NF-κB) and the release of antimicrobial cytokines, such as thymic stromal lymphopoietin (TSLP) [11][12][13] . TSLP, a four-helix bundle that function as an interleukin (IL)-17-like cytokine, is mostly expressed by epithelial cells in the airways and ocular tissues [14] .…”