Dynamic integration of CXCL8 or CCL20 signaling with IL-15 or IL-32α reprograms human T-cell functional plasticity

Tissue-derived chemokines can direct immune cell migration by engaging specific receptors, thereby controlling the localization and trafficking of immune cells within tissues. However, how chemokines modulate cytokines-driven transcriptional programs in T cells remain unclear. This study investigated how CXCL8 and CCL20 influence human T cell responses to IL-15 and IL-32α.

Peripheral blood T cells from health donors were evaluated for the expression and functional activity of the chemokine receptors CCR6, CXCR1, and CXCR2. Chemokine induced migration was assessed using in vitro migration assays. T cells were stimulated with CCL20 or CXCL8 alone or in combination with IL-15 or IL-32α, and transcriptional responses were characterized by transcriptomic profiling. Selected transcriptional findings were further evaluated through functional and protein-level assays.

Peripheral blood T cells expressed CCR6, CXCR1, and CXCR2, which were downregulated during vitro culture, but remained functionally responsive in migration assays, highlighting the influence of the microenvironmental on receptor expression. Transcriptomic profiling showed that CCL20 and CXCL8 independently reprogrammed GPCR-associated networks involved in migration, cytoskeletal dynamics, adhesion, and chemokine scavenging. IL-15 induced robust JAK/STAT-dependent inflammatory and survival programs together with CISH upregulation, indicating coordinated activation and negative-feedback regulation. In contrast, IL-32α modulated PI3K/AKT, adhesion, and NOTCH1-related pathways and was associated with increased T cell proliferation. Combined stimulation revealed context-dependent integration of these signaling programs. IL-15 plus CCL20 enhanced inflammatory and migratory signatures, whereas CXCL8 plus IL-15 produced a transcriptional profile largely dominated by IL-15. IL-32α plus CXCL8 induced cooperative programs involving vesicular remodeling, mTOR-associated metabolism, cytoskeletal organization, and cell-cycle regulation.

These findings demonstrated that chemokine and cytokine signals are integrated in a context-dependent manner to regulate T cells migration, activation, survival and proliferation. Rather than acting solely as chemoattractant, CCL20 and CXCL8 can reshape cytokine-driven transcriptional profile programs, revealing additional regulatory layer that may contribute to T cell functional plasticity within inflammatory and tumor microenvironment.