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

Inhal Toxicol. 2022;34(13-14):399-411. doi: 10.1080/08958378.2022.2134526. Epub 2022 Oct 19.

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; macrophages; scRNA-seq; t cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acute Lung Injury* / chemically induced
  • Acute Lung Injury* / genetics
  • Animals
  • Anti-Inflammatory Agents / pharmacology
  • Chlorine* / toxicity
  • Female
  • Lipopolysaccharides / toxicity
  • Lung / pathology
  • Macrophages
  • Mice
  • Mice, Inbred BALB C
  • Sequence Analysis, RNA
  • T-Lymphocytes

Substances

  • Chlorine
  • Anti-Inflammatory Agents
  • Lipopolysaccharides