Cigarette smoke sustains immunosuppressive microenvironment inducing M2 macrophage polarization and viability in lung cancer settings

PLoS One. 2024 May 22;19(5):e0303875. doi: 10.1371/journal.pone.0303875. eCollection 2024.

Abstract

Background: It is amply demonstrated that cigarette smoke (CS) has a high impact on lung tumor progression worsening lung cancer patient prognosis and response to therapies. Alteration of immune cell types and functions in smokers' lungs have been strictly related with smoke detrimental effects. However, the role of CS in dictating an inflammatory or immunosuppressive lung microenvironment still needs to be elucidated. Here, we investigated the effect of in vitro exposure to cigarette smoke extract (CSE) focusing on macrophages.

Methods: Immortalized murine macrophages RAW 264.7 cells were cultured in the presence of CS extract and their polarization has been assessed by Real-time PCR and cytofluorimetric analysis, viability has been assessed by SRB assay and 3D-cultures and activation by exposure to Poly(I:C). Moreover, interaction with Lewis lung carcinoma (LLC1) murine cell models in the presence of CS extract were analyzed by confocal microscopy.

Results: Obtained results indicate that CS induces macrophages polarization towards the M2 phenotype and M2-phenotype macrophages are resistant to the CS toxic activity. Moreover, CS impairs TLR3-mediated M2-M1 phenotype shift thus contributing to the M2 enrichment in lung smokers.

Conclusions: These findings indicate that, in lung cancer microenvironment of smokers, CS can contribute to the M2-phenotype macrophages prevalence by different mechanisms, ultimately, driving an anti-inflammatory, likely immunosuppressive, microenvironment in lung cancer smokers.

MeSH terms

  • Animals
  • Carcinoma, Lewis Lung / immunology
  • Carcinoma, Lewis Lung / pathology
  • Cell Polarity / drug effects
  • Cell Survival / drug effects
  • Humans
  • Lung Neoplasms* / pathology
  • Macrophage Activation / drug effects
  • Macrophages* / drug effects
  • Macrophages* / immunology
  • Macrophages* / metabolism
  • Mice
  • RAW 264.7 Cells
  • Smoke / adverse effects
  • Tumor Microenvironment* / drug effects

Substances

  • Smoke

Grants and funding

This work was supported by the AIRC (Associazione Italiana per la Ricerca sul Cancro (grant number 12162 to ET and 24718 to LS); the Fondazione Umberto Veronesi (Fellowship 2018 and 2019 to FB); and the Universita`degli Studi di Milano (Piano di Sostegno alla Ricerca 2017). The funding sources had no role in the study design; the collection, analysis, and interpretation of the data; the writing of the manuscript; or the decision to submit the manuscript for publication. There was no additional external funding received for this study.