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Intratumoral Three-Cell-Type Clusters Are a Conserved Feature of Endogenous Antitumor Immunity

  • Sheela R. Damle
  • , Jason A. Carter
  • , Kristin E. Goodsell
  • , Jose M.B. Pineda
  • , Lindsay K. Dickerson
  • , Xiuyun Jiang
  • , Jacqueline L. Mudd
  • , Thomas Walsh
  • , Heidi L. Kenerson
  • , Jack Cernak
  • , Sardar Shahmir B. Chauhan
  • , Emily Beirne
  • , Sujata Jana
  • , Amanda L. Koehne
  • , Kiran R. Vij
  • , Marianna B. Ruzinova
  • , Raymond Yeung
  • , Shreeram Akilesh
  • , Eric A. Collisson
  • , Ryan C. Fields
  • I. Nicholas Crispe, Venu G. Pillarisetty

Research output: Contribution to journalArticlepeer-review

Abstract

Effective antitumor immunity ultimately depends on the priming and activation of tumor-specific cytotoxic CD8+ T cells; however, the role of intratumoral cell–cell immune interactions remains incompletely understood. Recent work has revealed that the temporospatial co-localization of dendritic cells (DC), T helper (Th) cells, and cytotoxic T lymphocytes (CTL) within the tumor immune microenvironment following immune checkpoint blockade correlates with clinical response. In this study, we report the integration of more than 1 million spatially resolved single-cell profiles across six spatial proteomic and transcriptomic assays, which demonstrated that DC:Th:CTL three-cell-type clusters were common even in immunotherapy-naïve and highly desmoplastic tumors, such as fibrolamellar carcinoma and pancreatic ductal adenocarcinoma. We found that these immune triads were enriched for functionally important type 1 conventional DC, mature DCs enriched in immunoregulatory molecules, CXCL13+ Th, and GZMK+ effector CTL phenotypes. Subsequent multiplex immunofluorescence imaging of more than 450 primary pancreatic ductal adenocarcinoma tumors showed that the density of antigen-presenting cell:Th:CTL three-cell-type clusters was correlated with intratumoral T-cell clonal expansion and improved overall survival. These findings suggest that DC:Th:CTL triads are conserved across solid tumors and highlight the importance of intratumoral spatial niches in mediating endogenous antitumor immunity.

Original languageEnglish
Pages (from-to)205-218
Number of pages14
JournalCancer immunology research
Volume14
Issue number2
DOIs
StatePublished - Feb 1 2026

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