First-in-human single-cell and TCR atlas reveals peripheral immune remodeling before and after irreversible...

Prostate cancer (PCa) is the second most common malignancy in men worldwide. Although radical prostatectomy effectively controls tumor progression, it often causes complications such as urinary incontinence and sexual dysfunction. Irreversible electroporation (IRE), a non-thermal ablation technique that preserves neurovascular structures, has emerged as a promising focal therapy and may induce systemic antitumor immunity through immunogenic cell death. However, the mechanisms and temporal dynamics of systemic immune responses to IRE in humans remain unclear.

This prospective study enrolled five patients with localized PCa (T2a-T2c). Peripheral blood samples were collected before and 7 days after IRE treatment. Single-cell RNA sequencing and T-cell receptor (TCR) repertoire profiling (10x Genomics) were performed to construct a high-resolution immune landscape. Integrated bioinformatic analyses-including Seurat/Harmony clustering, differential expression, functional enrichment, Gene Set Variation Analysis, transcription factor analysis, cell-cell communication (CellChat), and clonal tracking (Immunarch/scRepertoire)-were used to characterize systemic immune modulation following IRE.

IRE induced bidirectional remodeling of the peripheral immune landscape. Myeloid cells, especially non-classical monocytes, increased in proportion and exhibited activation of inflammatory (interleukin-27), antiviral, complement, and antigen presentation pathways. T cells declined and exhibited reduced activation-associated and proliferation-associated transcriptional programs alongside enhanced survival-related signaling, consistent with a relative quiescent state of peripheral adaptive immunity at day 7. Natural killer cells showed elevated cytotoxic activity. TCR profiling revealed increased diversity and antigen-driven clonal expansion, while cell-cell communication shifted toward proinflammatory (TNF/SELPLG) and away from antigen-presenting (major histocompatibility complex (MHC)-I/CD40) interactions, indicating coordinated innate-adaptive rebalancing.

This study is the first to map systemic immune dynamics after IRE in prostate cancer using single-cell analysis. 1 week after IRE, peripheral blood mononuclear cells show robust innate activation with concomitant attenuation of circulating T-cell activation/proliferation and reduced MHC-I/CD40-related communication, consistent with a time-dependent systemic immune rebalancing phase rather than sustained peak adaptive activation in blood. These findings support time-resolved immune monitoring and may help optimize the timing of IRE-based combination immunotherapy.