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Tumour-reactive heterotypic CD8 T cell clusters from clinical samples.

Sofía Ibáñez-Molero1, Johanna Veldman1, Juan Simon Nieto1

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|November 19, 2025
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Summary

Researchers identified functional clusters of CD8+ T cells and tumor cells in melanoma. These T cell clusters, isolated from patient samples, show enhanced anti-tumor activity and potential for therapeutic development.

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Area of Science:

  • Immunology
  • Oncology
  • Cell Biology

Background:

  • CD8+ T cell proximity to tumor cells correlates with anti-tumor immunity.
  • The existence and functional significance of isolated T cell-tumor cell clusters are not well understood.

Purpose of the Study:

  • To investigate the presence and characteristics of heterotypic clusters of CD8+ T cells and tumor cells in human melanoma metastases.
  • To assess the functional potential and therapeutic utility of T cells isolated from these clusters.

Main Methods:

  • Conventional and imaging flow cytometry to isolate heterotypic clusters.
  • Single-cell RNA-sequencing to analyze T cell gene expression.
  • Ex vivo expansion and functional assays of T cells from clusters.
  • Adoptive cell transfer in mice to evaluate anti-tumor efficacy.

Main Results:

  • Heterotypic clusters of CD8+ T cells, tumor cells, and antigen-presenting cells (APCs) were isolated from all tested melanoma metastases.
  • T cells within clusters showed enrichment for tumor reactivity and exhaustion signatures, with increased TCR clonality.
  • Expanded T cells from clusters exhibited significantly enhanced killing activity against autologous melanomas and improved control in mouse models.
  • T cells from clusters demonstrated increased infiltration and activation in vivo.

Conclusions:

  • Tumor-reactive CD8+ T cells are enriched in functional, isolatable heterotypic clusters with tumor cells and/or APCs.
  • These T cell clusters represent a valuable source for understanding tumor-immune interactions and hold therapeutic potential.