Distinct Gene Expression Patterns between Nasal Mucosal Cells and Blood Collected from Allergic Rhinitis Sufferers

Abstract

Background: Investigations of gene expression in allergic rhinitis (AR) typically rely on invasive nasal biopsies (site of inflammation) or blood samples (systemic immunity) to obtain sufficient genetic material for analysis. New methodologies to circumvent the need for invasive sample collection offer promise to further the understanding of local immune mechanisms relevant in AR. Methods: A within-subject design was employed to compare immune gene expression profiles obtained from nasal washing/brushing and whole blood samples collected during peak pollen season. Twelve adults (age: 46.3 ± 12.3 years) with more than a 2-year history of AR and a confirmed grass pollen allergy participated in the study. Gene expression analysis was performed using a panel of 760 immune genes with the NanoString nCounter platform on nasal lavage/brushing cell lysates and compared to RNA extracted from blood. Results: A total of 355 genes were significantly differentially expressed between sample types (9.87 to –9.71 log₂ fold change). The top 3 genes significantly upregulated in nasal lysate samples were Mucin 1 (MUC1), Tight Junction Protein 1 (TJP1), and Lipocalin-2 (LCN2). The top 3 genes significantly upregulated in blood samples were cluster of differentiation 3e (CD3E), FYN Proto-Oncogene Src Family Tyrosine Kinase (FYN) and cluster of differentiation 3d (CD3D). Conclusions: Overall, the blood and nasal lavage samples showed vastly distinct gene expression profiles and functional gene pathways which reflect their anatomical and functional origins. Evaluating immune gene expression of the nasal mucosa in addition to blood samples may be beneficial in understanding AR pathophysiology and response to allergen challenge.

Adult allergic rhinitis sufferers have unique nasal mucosal and peripheral blood immune gene expression profiles: A case–control study

“…15 Nasal washing, brushing, and whole blood samples were collected as described previously. 12 Briefly, nasal wash samples were collected by instilling 100 ml of phosphate-buffered saline (PBS) in each nostril. Expelled fluid was collected in a sterile container and supplemented with 20 ml of RPMI media.…”

“… 11 We have successfully conducted gene expression experiments with cell lysate samples collected via nasal brushing and nasal washing using the NanoString nCounter system. 12 This method provided a cost‐effective and noninvasive means of sampling that yielded sufficient molecular material for expression analysis of a panel of 760 immune genes.…”

Beneficial effects of baicalein on a model of allergic rhinitis

“…During the late-phase of AR, other inflammatory cells are added, containing eosinophils, mast cells, basophils, and T cells, in the epithelium and the deeper lamina propria (22)(23)(24). Further release of histamine, leukotrienes, chemokines and other compounds maintains the anaphylactic reaction (6,25). In this study, the AR model of rats was established by repeated nasal drip OVA application, which exhibited similar allergic symptoms in humans; it can be used to analyze the detraction effect of AR (26).…”

Digital Immune Gene Expression Profiling Discriminates Allergic Rhinitis Responders from Non-Responders to Probiotic Supplementation

“…The assessment of immune differences between trial participants classified as “responders” and “non-responders” to probiotics may reveal key mucosal immune mechanisms underpinning the clinical effects of these supplements. New digital gene expression technology such as the Nanostring nCounter System is overcoming the limitation of accessing sufficient mucosal material to characterize the immunopathology of AR [5], and is an amplification-independent technique based on hybridization and direct digital counting that allows the simultaneous assessment of multiple targets for consideration of responses in the context of a biological network [6]. To examine the effect of probiotics on systemic and mucosal immunity, we utilized NanoString digital immune gene expression profiling in blood and nasal lavage/ nasal brushing samples in a subset of responders and non-responders from an eight-week supplementation study [4].…”