Transcriptomic profiling of human macrophages following adjuvant stimulation

Trained immunity is the phenomenon whereby innate immune cells such as monocytes or macrophages undergo functional reprogramming after exposure to certain microbial components, altering their responses to future exposures. Since trained immunity was first discovered, and has been commonly studied with the BCG vaccine, studies confirming trained immunity have often focused on human PBMCs. Thus, to study the mechanism of the vaccine in a well-established trained immunity model, we used human macrophages differentiated from monocytes isolated from PBMCs. The macrophages were expose to A + M + MA for 24 h, and rested two or five days prior to exposure to bacteria, as an ex vivo model with a five-day rest period has been used in numerous studies to confirm trained immunity. Overall design: PBMCs were obtained from healthy donors, and stimulated with the vaccine for 24 hours and rested for 2 or 5 days. Expeirment was repeated 3 times to obtain three biological replicates.