Abstract
Objective
Chlorine (Cl2), as an asphyxiant toxicant, induced poisoning incidents and acute lung injury (ALI) occur frequently. The specific pathogenesis of Cl2-induced ALI remains unclear. Immune cells play an important role in the process of lung damage. We used single-cell RNA sequencing (scRNA-seq) technology to explore T cells and macrophages molecular mechanism.
Methods
Female BALB/c mice were exposed to 400 ppm Cl2 for 15 min. scRNA-seq technology was used to observe the heterogeneity of T cells and macrophages. Hematoxylin–eosin (H&E) staining was used to evaluate the degree of lung injury. Immunofluorescence was used to verify the highly expressed genes of our interest.
Results
A total of 5316 to 7742 cells were classified into eight different cell types. Several new highly expressed anti-inflammatory and pro-inflammatory genes were found in T cells and macrophages, which were further verified in vitro. Through the pseudotime analysis of macrophages, it was found that the expression of pro-inflammatory and anti-inflammatory genes showed opposite trends in the development of Cl2-induced ALI. This study also mapped T cells-macrophage communication and identified the development of several important receptor-ligand complexes in Cl2-induced ALI.
Conclusions
These findings are worthy of further exploration and provide new resources and directions for the study of Cl2-induced ALI in mice, especially in immune and inflammation mechanisms.
Acknowledgements
The authors thank OE Biotech Co., Ltd (Shanghai, China) for providing single-cell RNA-seq.
Ethics approval and consent to participate
The experiment protocols were approved by the local animal ethics committee. All applicable international, national, and/or institutional guidelines for the care and use of animals.
Consent for publication
Not applicable.
Author contributions
CQ Zhao, JZ Liu and XD Zhang designed/performed most of the investigation, data analysis and wrote the manuscript; MM Liu and XT Ren provided pathological assistance; DQ Kong, J Peng and WH Yu contributed to interpretation of the data and analyses. All of the authors have read and approved the manuscript.
Disclosure statement
No potential conflict of interest was reported by the author(s).
Data availability statement
The datasets generated and analyzed during the present study are available from the corresponding author on reasonable request.