Mechanisms of interleukin 4 mediated increase in efficacy of vaccines against opioid use disorders.

We report our results of RNA-sequencing analysis of mouse lymph nodes and spleens after administration of an anti-oxycodone vaccine (OXY-sKLH) or carrier protein control (sKLH) with and without concurrent antibody-based depletion of interleukin-4 (IL-4) Overall design: Mice were vaccinated with OXY-sKLH or sKLH (control) with and without depletion of IL-4 using a monoclonal antibody. Lymph nodes and spleens were harvested on day 7 after vaccination and processed into a single cell suspension. Activated T cells were sorted via FACs and were defined as Ghost Dye- CD11c- F4/80- B220- CD8a- CD90.2+ CD4+ CD44+ cells. RNA was extracted from sorted cells using chloroform/trizol and collected using an RNA isolation kit. Samples were processed using a Clontech pico mammalian RNA library preparation. Libraries were sequenced on an Illumina NextSeq with 75 pair-end sequencing with samples acquiring an average of 30 million reads per sample.