Introduction: Olfactory dysfunction (OD) is a common problem affecting up to 20% of the general population. Previous studies identified olfactory cleft mucus proteins associated with OD in chronic rhinosinusitis (CRS) but not in a healthy population. This study aimed to identify olfactory cleft mucus proteins associated with olfaction in individuals without sinus disease. Methods: Subjects free of sinus disease completed medical history questionnaires collecting demographics, comorbidities and past exposures. Olfactory testing was performed using Sniffin' Sticks evaluating threshold, discrimination and identification. Olfactory cleft mucus (OC) and in select cases inferior turbinate mucus (IT) was collected with Leukosorb paper and assays performed for 17 proteins, including growth factors, cytokines/chemokines, cell cycle regulators and odorant binding protein (OBP). Results: A total of 56 subjects were enrolled with an average age of 47.8 years (SD 17.6 years), and included 33 of females (58.9%). Average Threshold/Discrimination/Identification (TDI) score was 30.3 (SD 6.4). In localization studies, OBP concentrations were significantly higher in OC than IT mucus (p=0.006). Cyclin-dependent kinase inhibitor 2A (CDKN2A/p16INK4a), basic fibroblast growth factor (bFGF), chemokine ligand 2 (CCL2/MCP-1), granulocyte macrophage colony-stimulating factor (GM-CSF), and chemokine ligand 20 (CCL20/MIP-3a) all inversely correlated with overall TDI (all rho≥−0.479, p≤0.004). Stem cell factor (SCF) positively correlated with overall TDI (rho=0.510, p=0.002).

Patients with CRSwNP and AFRS both have reduced sinonasal 1α-hydroxylase and 1,25VD3 compared to controls or CRSsNP. Reductions in intracellular 1α-hydroxylase combined from all sinonasal cell types were associated with more severe subjective disease severity in CRSwNP.

When HSNECs from CRSwNP patients are treated with SOD concurrently with Aspergillus or Alternaria, SOD treatment decreases the fungal antigen-induced inflammatory response. The ability to attenuate inflammation induced by common fungal allergens with SOD treatment could provide a novel therapeutic or preventative approach for patients with CRS or other allergic inflammatory airway diseases.

RationalePatients with chronic rhinosinusitis with nasal polyps (CRSwNP) have been shown to be vitamin D3 (VD3) deficient, which is associated with more severe disease and increased polyp size. To gain mechanistic insights into these observational studies, we examined the impact of VD3 deficiency on inflammation and VD3 metabolism in an Aspergillus fumigatus (Af) mouse model of chronic rhinosinusitis (Af-CRS).MethodsBalb/c mice were fed control or VD3 deficient diet for 4 weeks. Mice were then sensitized with intraperitoneal Af, and one week later given Af intranasally every three days for four weeks while being maintained on control or VD3 deficient diet. Airway function, sinonasal immune cell infiltrate and sinonasal VD3 metabolism profiles were then examined.ResultsMice with VD3 deficiency had increased Penh and sRaw values as compared to controls as well as exacerbated changes in sRaw when coupled with Af-CRS. As compared to controls, VD3 deficient and Af-CRS mice had reduced sinonasal 1α-hydroxylase and the active VD3 metabolite, 1,25(OH)2D3. Differential analysis of nasal lavage samples showed that VD3 deficiency alone and in combination with Af-CRS profoundly upregulated eosinophil, neutrophil and lymphocyte numbers. VD3 deficiency exacerbated increases in monocyte-derived dendritic cell (DC) associated with Af-CRS. Conversely, T-regulatory cells were decreased in both Af-CRS mice and VD3 deficient mice, though coupling VD3 deficiency with Af-CRS did not exacerbate CD4 or T-regulatory cells numbers. Lastly, VD3 deficiency had a modifying or exacerbating impact on nasal lavage levels of IFN-γ, IL-6, IL-10 and TNF-α, but had no impact on IL-17A.ConclusionsVD3 deficiency causes changes in sinonasal immunity, which in many ways mirrors the changes observed in Af-CRS mice, while selectively exacerbating inflammation. Furthermore, both VD3 deficiency and Af-CRS were associated with altered sinonasal VD3 metabolism causing reductions in local levels of the active VD3 metabolite, 1,25(OH)2D3, even with adequate circulating levels.

Background These studies examined the impact of environmental tobacco smoke (ETS) and active smoking on sinonasal dendritic cell (DCs) subsets in controls or patients with chronic rhinosinusitis with nasal polyps (CRSwNP). In subsequent in vitro investigations, we examined the influence of cigarette smoke extract (CSE) on human sinonasal epithelial cells’ (HSNECs) ability to regulate DC functions. Methods Sinonasal tissue, blood and hair were collected from patients undergoing sinus surgery. Smoking status and ETS exposure were determined by hair nicotine. DC subsets were examined by flow cytometric analysis. Monocyte-derived dendritic cells (moDCs) were treated with conditioned medium from non-smoked exposed HSNEC (NS-HSNEC) or cigarette smoke extract-exposed HSNEC (CSE-HSNEC) to assess the impact of CSE exposure on HSNEC regulation of moDCs functions. Results Control subjects who were active smokers displayed increased sinonasal moDC and mDC1 and reduced mDC2 cells, while in those with CRSwNP only moDC and mDC2 were altered. ETS was found to increase only moDCs in patients with CRSwNP. In vitro, CSE stimulated HSNEC secretion of the moDC regulatory products CCL20, PGE2 and GM-CSF. CSE-exposure also promoted HSNECs to stimulate monocyte and moDC migration. MoDCs treated with CSE-HSNEC media stimulated an increase in antigen uptake and expression of CD80 and CD86. Lastly, CSE-HSNEC treated moDCs secreted increased levels of IL-10, IFN-γ and TSLP compared Conclusions Active smoking and to a lesser degree ETS, alters the sinonasal composition of DCs. A potential mechanism to account for this is that cigarette smoke stimulates HSNEC to induce moDC migration, maturation and activation.