The development of T helper type 2 (Th2) responses in vivo is thought to require dendritic cells (DC) but how DCs initiate Th2 responses has remained elusive. We hypothesize that Th2 priming DC subsets are transcriptionally unique from DC that prime other inflammatory responses. We have probed the migratory DC subset transcriptomes of the Th2-driven mouse helminth infection model Heligmosomoides polygyrus bakerii (Hp) and Th17-driven mouse bacterial colitis infection model Citrobacter rodentium with direct ex vivo analysis to understand the differences between Th2 and Th17 priming DCs in the mesenteric lymph nodes (msLN).
RNA was prepared from sort-purified migratory CD11b+ and CD103+ DC subsets isolated from the msLN of C57BL/6 mice that were orally infected with either 200 stage 3 Hp larvae (8 days post infection) or 1-2 x 109 cfu Citrobacter rodentii (6 days post infection). RNA seq was performed by GeneWiz Inc using the Clontech SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing. Genome alignment was done with Tophat. EdgeR was used to generate FPKM values and perform statistical analysis. Real-time PCR was performed using Taqman probes. Flow cytometry was performed on a BD Canto. Cell sorting was performed on a BD Aria.
Comparing CD11b+ and CD103+ migratory DC from msLN of H polygyrus and Citrobacter rodentium infected wildtype mice, we identified 1267 and 667 differentially expressed genes, respectively.
Gene expression patterns of CD11b+ and CD103+ DC differ in H polygyrus and Citrobacter rodentium infections. We are further characterizing these differences through gene ontology and pathway analysis.