Heterogeneity of T cells and macrophages in chlorine-induced acute lung injury in mice using single-cell RNA sequencing

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.

Keywords:

• Chlorine
• acute lung injury
• scRNA-seq
• t cells
• macrophages

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.

Additional information

Funding

The work was supported by grants from the project of Military Medical Innovation [16CXZ021] & National Natural Science Foundation of China Youth Fund Project [No. 31900892].