ABSTRACT: Sleep insufficiency is associated with various disorders (cardiovascular disease, neurodegenerative diseases, immunological disorders.) but the molecular bases for these associations are largely unknown. We used multiple omics techniques to examine changes in the blood of healthy male and female adults voluntarily subjected to 24 h of sleep deprivation followed by up to one day of recovery with normal sleep routines. Thirty-two (14 males, 18 females) healthy adults 22-27 years old donated fasting blood samples prior to (Day 1) and following (Days 2 and 3) a 24-hour period of sleep deprivation. Blood samples were subjected to cell, biochemical, transcriptomic, proteomic and metabolomic analyses. Multi-omics integration analysis showed that sleep deprivation caused marked molecular changes (46.4% transcript genes, 59.3% proteins and 55.6% metabolites) that had not been restored by Day 2. Transient sleep deprivation had the greatest impact on the immune system and, in particular, on neutrophil-mediated immune processes. Correlation analysis with neutrophils suggested the plasma level of superoxidase dismustase-1 (SOD1) and the primary blood mononuclear cell transcriptional profile of the S100A8 gene could be used as biomarkers reflecting the sleep deprivation-associated disordered immunological state. An increased number of immune cells (leukocytes, neutrophils, monocytes, eosinophils, CD4+T), inflammatory factors (TNF-α, INF-γ, Il-6, IL-10, IL-12) and c-reactive protein in blood plasma further demonstrated the dysregulation of the immune system associated with short-term sleep deprivation. Other important findings associated with sleep deprivation included a significant decrease in the plasma melatonin level and an enrichment of molecules in signaling pathways associated with brain disorders, including schizophrenia and neurodegenerative diseases. Taken in concert, these points suggest that disrupted sleep, such as may occur in shift work, perturbs immunological status and may increase the risk for brain disorders in later life.