LAUSR

&NA; Agonists of nucleotide oligomerization domain (NOD) 1 and NOD2 receptors represent a promising class of immunostimulants and immunological adjuvants. Here, we describe a cell‐based test system to assess their pharmacokinetics. In this system, NOD1 and NOD2 agonist concentrations in sera are determined using a reporter cell line, 293Luc, which contains an NF‐&kgr;B‐inducible luciferase reporter construct and naturally expresses NOD1 and NOD2. The 293Luc cells dose‐dependently respond to different NOD1 and NOD2 agonists in the nanomolar to low‐micromolar concentration range. To verify that the NF‐&kgr;B‐inducing activity of serum samples is due to the administered agonist and not to secondarily induced endogenous molecules, a 293Luc‐derived NOD1/NOD2 double‐knockout clone is used. Within‐run and between‐run precisions of the system are <15% and <20%, respectively. Applicability of the novel assay is illustrated by studying pharmacokinetics of two specific NOD2 agonists (N‐acetyl‐d‐glucosaminyl‐N‐acetyl‐d‐muramyl‐l‐alanyl‐d‐isoglutamine and N‐glycolyl‐d‐muramyl‐l‐alanyl‐d‐isoglutamine) and a specific NOD1 agonist (N‐acetyl‐d‐glucosaminyl‐N‐acetyl‐d‐sorbitolamine‐d‐lactoyl‐l‐alanyl‐d‐isoglutamyl‐meso‐diaminopimelic acid). In summary, the test system described here can potentially be used to assess pharmacokinetics of NOD1 and NOD2 agonists in different animal species. HighlightsA cell‐based system to assess pharmacokinetics of NOD1 and NOD2 agonists is proposed.NOD1/NOD2 agonist concentrations in sera are measured by a 293Luc reporter cell line.293Luc cells dose‐dependently respond to nanomolar concentrations of the agonists.A 293Luc‐derived NOD1/NOD2 double‐knockout clone is used as a specificity control.Within‐run and between‐run precisions are <15% and <20%, respectively.