Functionally heterogeneous intratumoral CD4 <sup>+</sup> CD8 <sup>+</sup> double-positive T cells can give rise to single-positive T cells

T Tony Li (Department of Materials Science and Engineering, Stanford University) A Arielle Ilano (Division of Hematology/Oncology, Department of Medicine, University of California) M Marcel Arias-Badia (Division of Hematology/Oncology, Department of Medicine, University of California) D Diamond Luong (Division of Hematology/Oncology, Department of Medicine, University of California) H Hewitt Chang (Division of Hematology/Oncology, Department of Medicine, University of California) S Serena S. Kwek (Division of Hematology/Oncology, Department of Medicine, University of California) K Kathryn Allaire (Division of Hematology/Oncology, Department of Medicine, University of California) A Arun Chumber (Division of Hematology/Oncology, Department of Medicine, University of California) M Mason Sakamoto (Division of Hematology/Oncology, Department of Medicine, University of California) M Matthew Clark (Division of Hematology/Oncology, Department of Medicine, University of California) A Averey Lea (Division of Hematology/Oncology, Department of Medicine, University of California) M Mark Bridge (Helen Diller Family Comprehensive Cancer Center, University of California) B Brandon Chen E Eric Liu (Division of Hematology/Oncology, Department of Medicine, University of California) S Sima Porten (Division of Hematology/Oncology, Department of Medicine University of California‐San Francisco San Francisco California USA) M Maxwell V. Meng (Department of Urology, University of California) L Lauren I. R. Ehrlich (Department of Molecular Biosciences, The University of Texas at Austin) D David Y. Oh (Division of Hematology/Oncology, Department of Medicine, University of California) L Lawrence Fong (Division of Hematology/Oncology, Department of Medicine, University of California)

Abstract

Conventional single-positive (SP) CD4 + and CD8 + T cells recognize tumor antigens and help mediate clinical responses with cancer immunotherapy. Double-positive CD4 + CD8 + (DP) T cells have also been described in human cancers, but their role in the tumor microenvironment remains unclear. By generating a multiomic single cell atlas of DP and SP T cells, we find that DP T cells possess phenotypic heterogeneity similar to SP T cells that includes multiple clonally expanded populations of cytotoxic DP T cells in human renal cell carcinoma (RCC). These intratumoral DP T cells can mediate both MHC class I- and class II-dependent killing of autologous tumor cells. In addition, transcriptional profiling of DP TCR-bearing T cells revealed a gene signature enriched for clinical responders to PD-1 blockade in advanced RCC. We confirm prior observations of SP T cells transitioning into DP T cells and more notably, demonstrate that intratumoral T cells are capable of bidirectional differentiation in which DP T cells serve as precursors to SP T cell sin vivo. In the latter scenario, intratumoral DP T cells are shown to express Rag2 , suggesting that the tumor may act as an extrathymic site of T cell development. These findings reveal the multiple roles that DP T cells can possess in antitumor immunity.

Article Details

Volume / Issue Vol. 123, Issue 4
Published January 27, 2026
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (19)

T

Tony Li

Department of Materials Science and Engineering, Stanford University

A

Arielle Ilano

Division of Hematology/Oncology, Department of Medicine, University of California

M

Marcel Arias-Badia

Division of Hematology/Oncology, Department of Medicine, University of California

D

Diamond Luong

Division of Hematology/Oncology, Department of Medicine, University of California

H

Hewitt Chang

Division of Hematology/Oncology, Department of Medicine, University of California

S

Serena S. Kwek

Division of Hematology/Oncology, Department of Medicine, University of California

K

Kathryn Allaire

Division of Hematology/Oncology, Department of Medicine, University of California

A

Arun Chumber

Division of Hematology/Oncology, Department of Medicine, University of California

M

Mason Sakamoto

Division of Hematology/Oncology, Department of Medicine, University of California

M

Matthew Clark

Division of Hematology/Oncology, Department of Medicine, University of California

A

Averey Lea

Division of Hematology/Oncology, Department of Medicine, University of California

M

Mark Bridge

Helen Diller Family Comprehensive Cancer Center, University of California

B

Brandon Chen

E

Eric Liu

Division of Hematology/Oncology, Department of Medicine, University of California

S

Sima Porten

Division of Hematology/Oncology, Department of Medicine University of California‐San Francisco San Francisco California USA

M

Maxwell V. Meng

Department of Urology, University of California

L

Lauren I. R. Ehrlich

Department of Molecular Biosciences, The University of Texas at Austin

D

David Y. Oh

Division of Hematology/Oncology, Department of Medicine, University of California

L

Lawrence Fong

Division of Hematology/Oncology, Department of Medicine, University of California