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Visual analysis of multidimensional CyTOF data from allergic asthmatics pre and post allergen challenge

Wang, J.M., Bhakta, N., Ansel, M. et al.

Allergen-induced airway inflammation is a central feature of asthma. The ability to deeply phenotype cells with mass cytometry (CyTOF) can lead to new insights into the immune response after allergen challenge (AC). We have developed a 33 marker CyTOF-based panel and applied it to human bronchoalveolar lavage (BAL) and whole blood cells from 10 subjects with atopic asthma in a segmental AC study. Because the data generated by CyTOF are high dimensional, traditional bivariate analysis is complicated and limited to traditionally-defined cell types. Therefore, we applied the unbiased visualization method, viSNE. We report data from our initial subject before (baseline) and after diluent (DIL) or AC. Our results show that viSNE captures marker expression differences, cell grouping differences by sample type and rare cell populations. BAL cells showed more macrophages (CD206+) at baseline and more macrophages and neutrophils (CD16++CD15++CD24+) after DIL versus AC. In contrast, AC led to a tremendous influx of eosinophils (CD24++CCR3+) with some basophils (HLA-DRCD123+FcɛRI+). Interestingly, eosinophils in AC were CD69+ in the BAL but not in the blood, implying that eosinophils were activated after recruitment into the airways. There were no changes in T cell (CD3+CD4+ or CD3+CD8+), B cell (CD19+ or CD20+) or dendritic cell (CD1c+CD123HLA-DR+ or CD1cCD123+HLA-DR+) proportions between baseline, DIL and AC BAL cells; additional analyses from this clinical study will focus on subsets of these cells. In conclusion, visual analysis of high-dimensional data in allergic airway inflammation identifies differences and potentially meaningful but as yet undefined inflammatory cell populations.

Citation

Wang, J.M., Bhakta, N., Ansel, M. et al. "Visual analysis of multidimensional CyTOF data from allergic asthmatics pre and post allergen challenge" Journal of Immunology (2018): vol. 200 no. 1 Supplement 44.1