Project description:Segmental allergen challenge increases the percentage of eosinophils in bronchoalveolar lavage (BAL) cells. Mepolizumab, an anti-IL-5 therapeutic antibody, decreases the number of eosinophils in bronchoalveolar lavages (BAL). The use of both procedures allows to define genes that are either expressed by eosinophils or dependent on eosinophil presence in the airways.
Project description:Segmental allergen challenge increases the percentage of eosinophils in bronchoalveolar lavage (BAL) cells. Mepolizumab, an anti-IL-5 therapeutic antibody, decreases the number of eosinophils in bronchoalveolar lavages (BAL). The use of both procedures allows to define genes that are either expressed by eosinophils or dependent on eosinophil presence in the airways. Cells from Bronchoalveolar lavages (BAL) are obtained by bronchoscopy before (V5) and 48 h after a segmental allergen challenge (V6) in atopic and mild astmatics. This procedure was repeated in the same two subjects 2 months later and 1 month after an injection of mepolizumab (V22 is before challenge and V23 after challenge). Cells were immediately lysed and isolated total RNAs were analyzed using the Human Genome 1.0 ST GeneChip arrays (Affymetrix, Santa Clara, CA).
Project description:Genome wide DNA methylation profiling of 4 cell populations purified from paediatic bronchoalveolar lavage samples. The cell types profiled are alveolar macrophages, granulocytes, lymphocytes and alveolar epithelial cells. Fluorescence-activated single cell sorting was used to purify the cell populations of interest using previously described methods (Shanthikumar, AJRCMB, 2020 ;63(2):152-159). Genome wide DNA methylation profiling was performed via the EPICArray. These data can be used for reference based deconvolution of cell proportion of bronchoalveolar lavage samples. DNA extracted from raw BAL (n=6) was also profiled using the EPICArray, allowing for validation of reference based deconvulution.
Project description:Chronic beryllium disease (CBD) is a metal-induced hypersensitivity disorder characterized by CD4+ T cell alveolitis. Single-cell gene expression profiling of bronchoalveolar lavage (BAL) cells focused on T cells may provide insight into unique subsets of CD4+ T cells, functional pathways, and/or molecules that could potentially be developed into novel therapeutics for treating CBD and other granulomatous lung diseases. To understand the unbiased cellular composition of airway associated CD4+ T cells in subjects with beryllium exposure, we used BAL cells from one beryllium-exposed (BeS) and one CBD subject diagnosed using previously defined criteria, and these two samples comprised the disease cohort.