Optimizing sample collection to facilitate immune-signature of airway diseases by Mass cytometry
2Pulmonology Institute and CF Center, Carmel Medical Center, Israel
3B. Rappaport Faculty of Medicine, Technion - Israel Institute of Technology, Israel
The characterization of different inflammatory phenotypes of suppurative airway diseases such as PCD, CF, and bronchiectasis has important clinical implications. Investigating specific therapies (such as anti IL5 antibodies, and therapies targeted at neutrophilic activation) for specific phenotypes are dependent on inflammatory subtypes. A limited correlation has been found between sputum and blood inflammatory markers, reflecting the notion that inflammation may be uniquely confined to the airways in some diseases. To capture the uniqueness of the airway inflammatory cells, a mass cytometry approach was applied.
Mass Cytometry is among the powerful new technologies that had advanced the in-depth phenotyping capabilities, permitting high throughput quantification of over 40 parameters at single-cell resolution. However, due to differences in sample origin and cell subset of interest, specific experimental workflow needs to be developed in order to allow proper interpretation of the dynamic disease-associated changes in immune cell frequency or phenotype.
Here we optimized a sample process workflow to enable clinical sputum collection and preservation while maintaining protein staining using different approaches. Our workflow utilizes a 40-marker immune-phenotyping panel and SmartTube Proteomic Stabilizer, which allows for simple and reliable fixation and cryopreservation of sputum samples. We validate a workflow that allows collection of sputum samples with minimal processing requirements or expertise at the site of sample collection, followed by shipment to the CyTOF core facility for downstream staining and data acquisition.