Simple SummaryGenetic selection for immune response features may be a useful strategy for enhancing animal health. In this study, we identified the DEGs for dairy bulls with high- and low-AMIR. Results revealed that several DEGs were substantially associated with the regulation of locomotion, tissue development, immune response, and detoxification. In addition, the results of the KEGG pathway analysis showed that most DEGs were enriched in pathways related to disease, inflammation, and immune response, providing insights for genomic selection. These findings help to better comprehend the immune response mechanism in dairy bulls.AbstractEnhancing the immune response through breeding is regarded as an effective strategy for improving animal health, as dairy cattle identified as high immune responders are reported to have a decreased prevalence of economically significant diseases. The identification of differentially expressed genes (DEGs) associated with immune responses might be an effective tool for breeding healthy dairy cattle. In this study, antibody-mediated immune responses (AMIRs) were induced by the immunization of hen egg white lysozyme (HEWL) in six Chinese Holstein dairy bulls divided into high- and low-AMIR groups based on their HEWL antibody level. Then, RNA-seq was applied to explore the transcriptome of peripheral whole blood between the two comparison groups. As a result, several major upregulated and downregulated genes were identified and attributed to the regulation of locomotion, tissue development, immune response, and detoxification. In addition, the result of the KEGG pathway analysis revealed that most DEGs were enriched in pathways related to disease, inflammation, and immune response, including antigen processing and presentation, Staphylococcus aureus infection, intestinal immune network for IgA production, cytokine–cytokine receptor interaction, and complement and coagulation cascades. Moreover, six genes (BOLA-DQA5,C5,CXCL2,HBA,LTF, andCOL1A1) were validated using RT-qPCR, which may provide information for genomic selection in breeding programs. These results broaden the knowledge of the immune response mechanism in dairy bulls, which has strong implications for breeding cattle with an enhanced AMIR.