3D4 cells exhibit transcriptional features inconsistent with alveolar macrophage identity.

ABSTRACT: The 3D4 cell lines are widely used as porcine alveolar macrophage models in immunological research. However, our preliminary experiments revealed that 3D4/21 cells failed to respond to lipopolysaccharide stimulation. We challenged both 3D4/21 and 3D4/2 clones with various Toll-like receptor (TLR)4 (LPS, Ultrapure LPS, Kdo2-Lipid A) and TLR2/TLR1 (Pam3CSK4) ligands. Strikingly, all stimulants failed to activate pro-inflammatory responses (IL1B and TNF expression). To clarify the lack of response of 3D4 cells, we isolated primary porcine alveolar macrophages (PAM) from piglets using lung lavage, and analyzed the transcriptome of unstimulated or Kdo2-Lipid A-stimulated 3D4/21 cells compared with equally treated PAM. We identified extensive transcriptomic differences, with 10,718 differentiall

SUBMITTER: Ma W 

PROVIDER: S-EPMC12539023 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature