Single-cell gene expression analysis of cryopreserved equine bronchoalveolar cells

Frontiers in Immunology

Immunology

Laureen M. Peters¹  Vidhya Jagannathan²  Pamela Nicholson³  Tosso Leeb⁴  Sophie E. Sage⁵  Vinzenz Gerber⁵ 

[1] Clinical Diagnostic Laboratory, Department of Clinical Veterinary Medicine, Vetsuisse Faculty, University of Bern, Bern, Switzerland;Institute of Genetics, Vetsuisse Faculty, University of Bern, Bern, Switzerland;Next Generation Sequencing Platform, University of Bern, Bern, Switzerland;Next Generation Sequencing Platform, University of Bern, Bern, Switzerland;Institute of Genetics, Vetsuisse Faculty, University of Bern, Bern, Switzerland;Swiss Institute of Equine Medicine, Department of Clinical Veterinary Medicine, Vetsuisse Faculty, University of Bern, Bern, Switzerland;

关键词: single-cell mRNA sequencing;    cell cryopreservation;    equine respiratory system;    equine immunology;    cell annotation;   

DOI  :  10.3389/fimmu.2022.929922

 received in 2022-04-27, accepted in 2022-08-08,  发布年份 2022

来源: Frontiers

【 摘 要 】

The transcriptomic profile of a cell population can now be studied at the cellular level using single-cell mRNA sequencing (scRNA-seq). This novel technique provides the unprecedented opportunity to explore the cellular composition of the bronchoalveolar lavage fluid (BALF) of the horse, a species for which cell type markers are poorly described. Here, scRNA-seq technology was applied to cryopreserved equine BALF cells. Analysis of 4,631 cells isolated from three asthmatic horses in remission identified 16 cell clusters belonging to six major cell types: monocytes/macrophages, T cells, B/plasma cells, dendritic cells, neutrophils and mast cells. Higher resolution analysis of the constituents of the major immune cell populations allowed deep annotation of monocytes/macrophages, T cells and B/plasma cells. A significantly higher lymphocyte/macrophage ratio was detected with scRNA-seq compared to conventional cytological differential cell count. For the first time in horses, we detected a transcriptomic signature consistent with monocyte-lymphocyte complexes. Our findings indicate that scRNA-seq technology is applicable to cryopreserved equine BALF cells, allowing the identification of its major (cytologically differentiated) populations as well as previously unexplored T cell and macrophage subpopulations. Single-cell gene expression analysis has the potential to facilitate understanding of the immunological mechanisms at play in respiratory disorders of the horse, such as equine asthma.

【 授权许可】

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Copyright © 2022 Sage, Nicholson, Peters, Leeb, Jagannathan and Gerber