Cryopreserved Equine Bronchoalveolar Cell Single-Cell Gene Expression Analysis
Summary
Amid the prevalent immunosuppression observed in septic patients, the broader landscape of the immune response to sepsis remains under-explored. Their study utilized single-cell RNA sequencing on a CLP mouse model, revealing considerable variations in canonical immune cells during sepsis across key tissue sites. One significant finding was the identification of a unique dendritic cell subcluster—termed mregDCs—akin to those described in cancer research. These mregDCs were found to strongly engage naïve CD4+ T cells and encourage their transformation into regulatory T cells. This phenomenon seems linked to specific signaling pathways activated within the first day of septic challenge. Furthermore, parallels were drawn with human sepsis data from a recent COVID-19 patient study.
Research Criteria
These horses suffered from mild-to-moderate equine asthma but were in clinical remission at the time of the study. No medication other than alpha-2 agonists (for sedation) was administered for at least a month before the experiment. The horses' lower airway inflammation status was assessed via clinical scoring, bronchoscopy, and bronchoalveolar lavage fluid (BALF) cytology.
Fig.1 Experimental design1.
Sample Type
The article utilized single-cell mRNA sequencing (scRNA-seq) to study the transcriptomic profile of cell populations at the cellular level.
Result—Cell Distribution
Three university-owned horses diagnosed with mild-to-moderate equine asthma were studied. Despite their asthma, they were deemed healthy overall. The diagnosis was based on specific clinical and cytological criteria, and at the time of the study, the horses were in clinical remission. A single-cell RNA sequencing (scRNA-seq) was conducted on 4,631 cells from these horses, revealing 16 distinct clusters. These clusters were grouped into six major cell populations, including monocytes/macrophages, dendritic cells, T cells, B/Plasma cells, neutrophils, and mast cells. The distribution of these cell types was consistent across the three horses, with a notable exception being a higher proportion of mast cells in one horse. Cryopreservation, a process used in the study, did not significantly alter the distribution of cell types. The scRNA-seq results showed a higher lymphocyte/macrophage ratio compared to traditional cytology.
Fig.2 Analysis of 4,631 cryopreserved equine bronchoalveolar cells obtained from three horses using scRNA-seq1.
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Reference
- Sage, Sophie E., et al. "Single-cell gene expression analysis of cryopreserved equine bronchoalveolar cells." Frontiers in immunology (2022): 4877.
