γδ T-cell Receptors Derived from Breast Cancer–Infiltrating T Lymphocytes Mediate Antitumor Reactivity

Abstract

γδ T cells in human solid tumors remain poorly defined. Here, we describe molecular and functional analyses of T-cell receptors (TCR) from tumor-infiltrating γδ T lymphocytes (γδ TIL) that were in direct contact with tumor cells in breast cancer lesions from archival material. We observed that the majority of γδ TILs harbored a proinflammatory phenotype and only a minority associated with the expression of IL17. We characterized TCRγ or TCRδ chains of γδ TILs and observed a higher proportion of Vδ2+ T cells compared with other tumor types. By reconstructing matched Vδ2– TCRγ and TCRδ pairs derived from single-cell sequencing, our data suggest that γδ TILs could be active against breast cancer and other tumor types. The reactivity pattern against tumor cells depended on both the TCRγ and TCRδ chains and was independent of additional costimulation through other innate immune receptors. We conclude that γδ TILs can mediate tumor reactivity through their individual γδ TCR pairs and that engineered T cells expressing TCRγ and δ chains derived from γδ TILs display potent antitumor reactivity against different cancer cell types and, thus, may be a valuable tool for engineering immune cells for adoptive cell therapies.

Aberrantly Expressed Embryonic Protein NODAL Alters Breast Cancer Cell Susceptibility to γδ T Cell Cytotoxicity

“…In contrast, the activity of γδ T cell TIL in TNBC in situ has been painstakingly investigated in a recently published study in which γδ T cells were identified in frozen TNBC tumor sections from nine patients, isolated by laser capture microdissection and subjected to single cell sequencing analysis. These analyses confirmed a polyclonal population of γδ T cells had infiltrated TNBC tumors and that these expressed CD69 and the proinflammatory cytokines IFNγ and TNFα; only a minor fraction (<20%) expressed IL-17 (30). Since different combinations of TCRγ and TCRδ chains confer distinct antigen recognition capabilities (30), if the response of γδ T cells to NODAL stimulation is TCR dependent, effects on individual clones would have been lost in our current analyses.…”

“…These analyses confirmed a polyclonal population of γδ T cells had infiltrated TNBC tumors and that these expressed CD69 and the proinflammatory cytokines IFNγ and TNFα; only a minor fraction (<20%) expressed IL-17 (30). Since different combinations of TCRγ and TCRδ chains confer distinct antigen recognition capabilities (30), if the response of γδ T cells to NODAL stimulation is TCR dependent, effects on individual clones would have been lost in our current analyses. As such, a study of the impact of NODAL on clonal populations may be of interest, or single cell RNAseq (39) could be employed to tease out individual responses.…”

“…Later studies have delved more deeply into the presence of γδ T cells infiltrating triple negative breast cancers (TNBC), revealing increased presence of γδ T cells compared to fibroadenomas or breast tissues from healthy individuals, suggesting active infiltration of γδ T cells into tumors (29), and that infiltrating γδ T cells are likely active (30).…”

Cancer immunotherapy with γδ T cells: many paths ahead of us

“…Wu et al detected high proportions of IL-17-producing cells among γδ TILs in colorectal cancer, 84 whereas Meraviglia et al reported only low numbers in a different cohort of colorectal cancer patients. 68 Very few IL-17-expressing γδ T cells among breast-cancer-infiltrating γδ TILs were reported in a recent study by Janssen et al 87 Again, we expect that better conclusions can be drawn in future studies using automated high-content imaging of tumor tissue to more precisely define and quantify immune cell composition within the tumor and peritumoral tissue. As yet, limited information is available on the TCR repertoire of γδ TILs in human tumors.…”

Adoptive Cell Therapy in Breast Cancer: A Current Perspective of Next-Generation Medicine

“…TILs compartment. gdT cells exhibit potent anti-tumor responses by bridging innate and adaptive immunities, since they incorporate both gdTCRs and killer cell immunoglobulin-like receptors (KIRs) (39,40). Also, gdT cell ligand recognition requires the expression of accessory costimulatory molecules, which may prevent harmful selfreactivity.…”