Bovine Herpesvirus 1 (BoHV1) is a leading cause of Bovine Respiratory Disease (BRD) in young calves, which is responsible for substantial morbidity and mortality. Therefore, the objective of the current study was to elucidate the mRNA transcriptomic response in mediastinal lymph node tissue to an experimental challenge with BoHV1, in dairy calves. Holstein-Friesian calves were either challenged by intranasal atomisation with BoHV1 virus (6.3 x 10^7/mL x 1.35mL) (n=12) or mock challenged with sterile phosphate buffered saline (n=6). Clinical signs were scored daily until euthanasia at day 6 post-challenge. Total RNA was extracted and sequenced from mediastinal lymph node tissue (100 bp paired-end). Sequence reads were aligned to the ARS-UCD1.2 bovine reference genome and differential gene expression analysis was performed using EdgeR. Anlysis uncovered a total of 81 differentially expressed genes (p < 0.05, FDR < 0.1, fold change > 2) between the BoHV-1 challenged and control calves. Overall design: Examination of a single tissue type (mediastinal lymph node) in two different treatment groups, BoHV-1 challenged calves (n=12) and control calves (n=6)