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Updated: Jan 17, 2026
Isolating Human Peripheral Blood Mononuclear Cells and CD4+ T cells from Sézary Syndrome Patients for Transcriptomic Profiling
Published on: October 14, 2021
Blood Transcriptome Analysis Reveals CTSB and ATP6V0D1 Expression in Circulating Monocytes as Potential Biomarkers of
Yaojun Peng1,2, Qiyan Wu3,4, Baimei Zhuang1
1Department of Emergency, The First Medical Center of Chinese PLA General Hospital, Beijing, China.
Background:
Sepsis is a dysregulated host response to infections, leading to organ dysfunction and posing a critical threat to human health. Despite tremendous progress in understanding the pathophysiology of sepsis, early diagnosis and clinical treatment efficacy remain unsatisfactory. This study aimed to identify transcriptomic alterations in peripheral blood mononuclear cells as potential biomarkers of sepsis.
Methods:
Bulk RNA-seq was performed on peripheral blood mononuclear cells obtained from 20 patients with sepsis and 12 healthy individuals. Multiple bioinformatics tools were used to identify key genes and signaling pathways associated with sepsis progression. The hub genes were further externally validated by publicly available blood transcriptomic data and experimentally verified by immunocytofluorescence assay.
Results:
Differential expression analysis revealed 4,522 differentially expressed genes in patients with sepsis (n = 20) compared with healthy individuals (n = 12). Weighted gene coexpression network analysis identified multiple gene modules closely related to sepsis, with the royal blue module exhibiting the most positive correlation with sepsis. Intersection analysis yielded 176 common genes between the royal blue module genes and differentially expressed genes. Protein-protein interaction analysis revealed five hub genes ( CTSB , CTSD , ATP6V0D1 , UBE2D1 , and ATP6V0C ) associated with sepsis. Immune infiltration was dissected by single-sample gene set enrichment analysis, revealing associations between hub genes and monocytes. Single-cell RNA sequencing data analysis and immunocytofluorescence assay confirmed the upregulation of CTSB and ATP6V0D1 in circulating monocytes. Notably, CTSB and ATP6V0D1 were significantly associated with 28-day mortality of sepsis patients in the external validation cohort (n = 479).
Conclusion:
This study identifies CTSB and ATP6V0D1 expression in circulating monocytes as potential biomarkers and promising therapeutic targets for sepsis.
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