Adaptive restoration of T-cell motility by cold acclimation through metabolic and transcriptional remodeling

Low temperatures reduce T-cell motility; however, the mechanisms by which T cells adapt to low-temperature environments remain poorly understood. Here, we investigated how T cells respond and adapt to subphysiological temperatures.

Conalbumin-specific D10 T cells were analyzed by time-lapse live-cell imaging using an inverted microscope equipped with a temperature-controlled chamber. Cellular respiration was assessed using a Seahorse XFe96 analyzer, and transcriptomic changes were examined by RNA sequencing.

At 37°C, T cells exhibited active lamellipodia-driven migration. Reducing the temperature below 32°C progressively suppressed crawling motility, accompanied by increased cell rounding. At 26°C, motility was markedly reduced in non-acclimated T cells. Unexpectedly, T cells transiently exposed to low temperatures during routine passaging retained motility under cold conditions, suggesting the induction of cold acclimation. Consistent with this interpretation, T cells cultured at temperatures between 30°C and 26°C for 24 h exhibited enhanced motility when evaluated at the corresponding temperatures compared with non-acclimated cells, with the most pronounced functional difference observed at 26°C. Cold acclimation at 26°C was also associated with a marked increase in mitochondrial spare respiratory capacity. RNA sequencing revealed extensive transcriptional reprogramming following cold acclimation, characterized by upregulation of pathways related to ribosome biogenesis, RNA processing, transcriptional regulation, and protein quality control, and downregulation of gene sets involved in DNA replication, DNA repair, and cell cycle progression, consistent with adaptive cellular remodeling.

These findings demonstrate that T-cell motility is highly temperature-sensitive, but prior exposure to low temperature enables adaptive restoration of migratory activity through coordinated metabolic and transcriptional remodeling. This adaptive response suggests that temperature actively regulates immune cell dynamics rather than serving solely as an environmental constraint and provides new insight into temperature-dependent immune regulation.