Novel Analysis of Immune Cells from Nasal Microbiopsy Demonstrates Reliable, Reproducible Data for Immune Populations, and Superior Cytokine Detection Compared to Nasal Wash

Abstract

The morbidity and mortality related to respiratory tract diseases is enormous, with hundreds of millions of individuals afflicted and four million people dying each year. Understanding the immunological processes in the mucosa that govern outcome following pathogenic encounter could lead to novel therapies. There is a need to study responses at mucosal surfaces in humans for two reasons: (i) Immunological findings in mice, or other animals, often fail to translate to humans. (ii) Compartmentalization of the immune system dictates a need to study sites where pathogens reside. In this manuscript, we describe two novel non-invasive nasal mucosal microsampling techniques and their use for measuring immunological parameters: 1) using nasal curettes to collect cells from the inferior turbinate and; 2) absorptive matrices to collect nasal lining fluid. Both techniques were well tolerated and yielded reproducible and robust data. We demonstrated differences in immune populations and activation state in nasal mucosa compared to blood as well as compared to nasopharyngeal lumen in healthy adults. We also found superior cytokine detection with absorptive matrices compared to nasal wash. These techniques are promising new tools that will facilitate studies of the immunological signatures underlying susceptibility and resistance to respiratory infections.

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

“…These results may reflect the compartmentalised nature of the immune system, where circulating blood and the nasal mucosa have a different cellular composition and biological roles. Jochems et al [10] have previ- ously compared immune cell populations and cytokine profiles in blood samples to nasal mucosal samples from healthy adults and reported that immune cells collected from the nose differed in composition and activation state than those in blood. In AR sufferers, nasal washing and brushing samples typically yield epithelial cells along with infiltrating immune cells such as eosinophils, neutrophils, and basophils [11][12][13].…”

“…In AR sufferers, nasal washing and brushing samples typically yield epithelial cells along with infiltrating immune cells such as eosinophils, neutrophils, and basophils [11][12][13]. In contrast to nasal mucosal samples, blood samples to do not contain epithelial cells and are more abundant in natural killer cells and monocytes [10,14]. Compared to the heat map analysis, additional insight into the pathogenesis of AR was gained when highly expressed genes were grouped into functional pathways.…”

Streptococcus pneumoniae: transmission, colonization and invasion

“…In mouse models, acquisition of the organism leads to a mild acute inflammatory response in the URT that is ineffective at completely clearing the organism 67 . By contrast, preexisting inflammation is the most closely associated susceptibility factor in the human challenge model 68 . Many of the factors contributing to the eventual clearance of S. pneumoniae have been delineated.…”

“…Pneumococcal biofilm structures have been detected in biopsy samples from patients with otitis media 109 and in the middle ear cleft of chinchillas co-infected with S. pneumoniae and Haemophilus influenzae 110 . In biopsy samples from volunteers colonized in experimental human studies, S. pneumoniae was found in microcolonies 68 , although it has not been determined whether these have characteristics of biofilms, such as an extracellular matrix. The production of an extracellular matrix has a major impact on the ability of S. pneumoniae that has been grown in a biofilm in vitro to subsequently translocate from the nasopharynx to the lungs in a murine infection model 111 .…”

Safety of Cultivated Limbal Epithelial Stem Cell Transplantation for Human Corneal Regeneration

“…The serum is harvested from bovine foetuses taken from pregnant cows during slaughter [47]. Irradiation of FBS is frequently used to eliminate live virus, but no process can be guaranteed as fully effective over time [48].…”