(P. aeruginosa cluster (green rods) and S. aureus (red spheres))
Respiratory failure due to acute and chronic infections and aberrant inflammatory responses, is the primary cause of death in patients with Cystic Fibrosis (CF). The significant expansion of infection and immunity related research in our CF Center prompted the establishment of a core, the Infection and Immunity (I&I) Core. The collective expertise of the I&I Core Director and Associate Directors sets the stage for innovative studies to examine defects in host response and microbial adaptation during CF, as well as testing new therapeutic approaches to tackle the destructive and life-threatening infections and inappropriate immune responses that play a major role in CF pathophysiology.
Goal: Provide experimental expertise, reagents and consultation in experimentation and preliminary data generation pertaining to immunology projects in CF research
Our six objectives:
Develop models to study host-pathogen interactions in CF
To support the CF Investigators with expertise to examine host-pathogen interactions in CF, we have developed assays to examine the interaction and colonization of CF pathogens with the respiratory epithelium, utilizing primary human respiratory epithelial cells from Core A and organotypic cultures, as well as CF clinical isolates of bacteria from Core D.
Provide immunological assays to assess dysfunction in CF
This core component will provide a comprehensive suite of immunologic assays to assess immune defects in CF, including multi-plex assays using the BioRad MagPix platform (which allows up to 50 cytokines to be analyzed at the same time), classical cytokine detection and quantification, flow cytometry, and enzymatic assays to assess immune cell function. Recently we have expanded the use of spectral flow cytometry, using the Cytek Aurora platform, allowing for complex immunophenotyping of human and mouse samples.
Provide animal models of lung infection, including epithelial-specific knockout models
Mouse models of acute and chronic infections with a variety of CF pathogens (bacterial, viral and fungal) have been developed, as well as polymicrobial infection models, histological assessments of lung injury and physiologic measurements, using the forced-oscillation technique.
Provide support and expertise for next-generation sequencing assays
To explore novel pathways involved in CF pathogenesis, the I&I Core will provide state-of-the-art next-gen sequencing methods including 16S sequencing, total RNA-seq, Dual-seq, ATAC-seq, single-cell RNA-seq, IgSeq, and spatial transcriptomics. Utilizing these methods will set our CF Investigators on pace to tackle pioneering microbiology and immunology questions.
Provide bioinformatics support
A crucial component to providing access to the latest next-gen sequencing technologies is bioinformatics and biostatistics consultation and analytical support, as well as expertise in predictive modeling, which will allow our teams to integrate sequencing data with patient metadata to identify new targets for therapeutic intervention to improve the disease course for CF patients.
Provide novel methods for gut to lung immune interaction studies
CF disease is characterized by multi-organ dysfunction. In the era of CFTR potentiator and corrector therapies, a renewed interest has been forged in the impact of CF on organs below the diaphragm. To address this increased focus, we have added expertise in sampling the lymphatic circulation that can link host-pathogen and immune interactions in the gut with the lung. This expertise will allow for studies focused on immunometabolism.