NKp46-expressing human gut-resident intraepithelial Vδ1 T cell subpopulation exhibits high antitumor activity against colorectal cancer
Joanna Mikulak, … , Antonino Spinelli, Domenico Mavilio
Joanna Mikulak, … , Antonino Spinelli, Domenico Mavilio
Published November 5, 2019
Citation Information: JCI Insight. 2019;4(24):e125884. https://doi.org/10.1172/jci.insight.125884.
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Research Article Gastroenterology Immunology

NKp46-expressing human gut-resident intraepithelial Vδ1 T cell subpopulation exhibits high antitumor activity against colorectal cancer

Abstract

γδ T cells account for a large fraction of human intestinal intraepithelial lymphocytes (IELs) endowed with potent antitumor activities. However, little is known about their origin, phenotype, and clinical relevance in colorectal cancer (CRC). To determine γδ IEL gut specificity, homing, and functions, γδ T cells were purified from human healthy blood, lymph nodes, liver, skin, and intestine, either disease-free, affected by CRC, or generated from thymic precursors. The constitutive expression of NKp46 specifically identifies a subset of cytotoxic Vδ1 T cells representing the largest fraction of gut-resident IELs. The ontogeny and gut-tropism of NKp46+/Vδ1 IELs depends both on distinctive features of Vδ1 thymic precursors and gut-environmental factors. Either the constitutive presence of NKp46 on tissue-resident Vδ1 intestinal IELs or its induced expression on IL-2/IL-15–activated Vδ1 thymocytes are associated with antitumor functions. Higher frequencies of NKp46+/Vδ1 IELs in tumor-free specimens from CRC patients correlate with a lower risk of developing metastatic III/IV disease stages. Additionally, our in vitro settings reproducing CRC tumor microenvironment inhibited the expansion of NKp46+/Vδ1 cells from activated thymic precursors. These results parallel the very low frequencies of NKp46+/Vδ1 IELs able to infiltrate CRC, thus providing insights to either follow-up cancer progression or to develop adoptive cellular therapies.

Authors

Joanna Mikulak, Ferdinando Oriolo, Elena Bruni, Alessandra Roberto, Federico S. Colombo, Anna Villa, Marita Bosticardo, Ileana Bortolomai, Elena Lo Presti, Serena Meraviglia, Francesco Dieli, Stefania Vetrano, Silvio Danese, Silvia Della Bella, Michele M. Carvello, Matteo Sacchi, Giovanni Cugini, Giovanni Colombo, Marco Klinger, Paola Spaggiari, Massimo Roncalli, Immo Prinz, Sarina Ravens, Biagio di Lorenzo, Emanuela Marcenaro, Bruno Silva-Santos, Antonino Spinelli, Domenico Mavilio

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Figure 6

Phenotype and functions of human of NKp46+ γδ thymocyte precursors following activations with physiologic and pathologic stimuli.

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Phenotype and functions of human of NKp46+ γδ thymocyte precursors follo...
(A) Summary statistical graphs showing the expression of CD4, CD8, CD56, NKG2A, NKG2D, CD16, and CCR9 on γδ thymocyte precursors (n ≥ 8) cultured either in the absence (Ctrl) or presence of IL-2. (B) Summary statistical graphs showing the CCL25 dose-dependent chemotaxis of FACS-sorted CCR9+/NKp46+ Vδ1 T cells generated from thymocyte precursors cultured with IL-2 for 6 days (n = 8). (C) Summary statistical graphs showing the transcripts levels of GZMB and NKp46 transcripts on freshly FACS-sorted thymocytes (Ctrl) and on NKp46+ and NKp46– Vδ1 T cells generated from thymocyte precursors cultured with IL-2 for 6 days (n = 6). Results are represented as the fold change (2–ΔΔCT) of target gene relative to a Ctrl samples and normalized to housekeeping genes GAPDH and S18. (D) Summary statistical graphs showing the expression of CD107a on NKp46+ and NKp46– Vδ1 T cells generated from thymocyte precursors cultured with IL-2 for 6 days either in the absence (Ctrl) or in the presence of K562 or Caco2 human tumor target cells (n ≥ 9). (E) Summary statistical graphs showing the NKp46 expression on freshly purified thymocytes (day 0) and on γδ thymocytes cultured with IL-2 for 6 days either in the absence (Ctrl) or in the presence of primary human colonic epithelial cells (HCoEpic) or IL-15, TGF-β, or Caco2 cell lines (n ≥ 8). *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001.