B cell-stromal cell cross talk drives mesenteric lymph node eosinophilia during intestinal helminth infection
We hypothesised that eosinophil activation and gene expression were linked to their interaction with the activated stroma within the lymph node. However, the role of eosinophil stromal crosstalk and how this modulate eosinophil function is unknown. To directly assess this hypothesis, we activated lymph node derived stromal cells using an agonist antibody for LT?R (Clone 4H8WH2). Considering eosinophils have long been associated with providing B cell proliferative and survival factors, we mimicked a cell-based activation model using purified naïve B cells given that B cell interactions with stroma can produce factors that can regulate eosinophil activation and migration. B cell-based activation as well as agonist antibody-based activation resulted in an activated stromal phenotype and were later used for co-culture with bone marrow-derived eosinophils. After 8 hours of co-culture with activated stroma, the eosinophils were purified, and bulk RNA sequencing was performed to identify global changes in their transcriptomes. Overall design: To investigate the impact of stromal cells on eosinophils gene expression. Bone marrow-derived eosinophils were generated in vitro and used for the co-culture experiment. Experiments were performed in triplicate wells and eosinophils were pooled post stimulation to create a single sample. RNAseq was performed using BGI Genomics DNBSEQ sequencing technology.