Abstract
Pathogenic chlamydial species restrict their peptidoglycan (PG) to the division septum of their replicative forms. PG is a microbe-associated molecular pattern (MAMP) and two of its major pattern recognition receptors in human cells are nucleotide-binding oligomerization domain-containing proteins 1 and 2 (NOD1 and NOD2, respectively). It has been proposed that this unique morphological feature is evidence of pathoadaptation by the microbe, permitting PG-dependent cell division while also reducing the bacterium's recognition by innate immune receptors. Chlamydia trachomatis-infected cells activate NOD1 signaling within 8-12 hours of exposure to the bacterium, roughly coinciding with the microbe's transition from its infectious to replicative forms. Here we report that, unlike NOD1 signaling, Chlamydia-induced NOD2 signaling does not occur until later in the pathogen's developmental cycle. Both C. trachomatis and the related murine pathogen Chlamydia muridarum signal late in infection in HEK293 reporter cell lines expressing either human or murinederived NOD2 receptors. NOD2 signaling can be modulated by disruption of the chlamydial amidase enzyme, AmiA(CT), interrupting the microbe's developmental cycle, and inducing RB lysis with inhibitors of lipooligosaccharide or peptidoglycan biosynthesis / assembly. These results mirror prior observations with Chlamydia-induced TLR9 signaling, leading us to hypothesize that Chlamydia-induced NOD2 signaling results from RB lytic events that occur sporadically during the RB to EB transition. Given our finding that pre-treating cells with NOD2-stimulatory ligands reduces chlamydial inclusion size, we hypothesize that the microbe preferentially degrades its PG during development to reduce the generation of NOD2 ligands, at the cost of enhancing NOD1 signaling.