Comparative analysis of methods to reduce activation signature gene expression in PBMCs

作者：Lovatiana Andriamboavonjy, Adam MacDonald, Laura K. Hamilton, Marjorie Labrecque, Marie‐Noëlle Boivin, Jason Karamchandani, Jo Anne Stratton, Martine Tétreault · 发表于：Scientific Reports · 年份：2023 · DOI：10.1038/s41598-023-49611-2 · 被引用次数：18 · 研究领域：RNA Interference and Gene Delivery、Immune Response and Inflammation、Immune Cell Function and Interaction

Preserving the in vivo cell transcriptome is essential for accurate profiling, yet factors during cell isolation including time ex vivo and temperature induce artifactual gene expression, particularly in stress-responsive immune cells. In this study, we investigated two methods to mitigate ex vivo activation signature gene (ASG) expression in peripheral blood mononuclear cells (PBMCs): transcription and translation inhibitors (TTis) and cold temperatures during isolation. Comparative analysis of PBMCs isolated with TTis revealed reduced ASG expression. However, TTi treatment impaired responsiveness to LPS stimulation in subsequent in vitro experiments. In contrast, cold isolation methods also prevented ASG expression; up to a point where the addition of TTis during cold isolation offered minimal additional advantage. These findings highlight the importance of considering the advantages and drawbacks of different isolation methods to ensure accurate interpretation of PBMC transcriptomic profiles.