TLR4 Activation Assay with NETs and Anti-CarP Antibodies

This experiment quantified Toll-like receptor 4 (TLR4) activation using a HEK293 reporter cell line to assess how NETs exposed to anti-carbamylated protein (anti-CarP) antibodies affect innate immune signaling. The study tested whether immune complexes formed between NETs and anti-CarP antibodies could enhance TLR4-mediated inflammatory responses. HEK293 cells engineered to express TLR4 and reporter genes were exposed to NETs that had been pre-incubated with or without anti-CarP IgG, and TLR4 activation was measured through reporter gene expression.