Cancer-associated fibroblast suppresses killing activity of natural killer cells through downregulation of poliovirus receptor (PVR/CD155), a ligand of activating NK receptor

Inoue,Tomoko; Adachi,Katsuyuki; Kawana,Kei; Taguchi,Ayumi; Nagamatsu,Takeshi; Fujimoto,Asaha; Tomio,Kensuke; Yamashita,Aki; Eguchi,Satoko; Nishida,Haruka; Nakamura,Hiroe; Sato,Masakazu; Yoshida,Mitsuyo; Arimoto,Takahide; Wada-Hiraike,Osamu; Oda,Katsutoshi; Osuga,Yutaka; Fujii,Tomoyuki

doi:10.3892/ijo.2016.3631

Cancer-associated fibroblast suppresses killing activity of natural killer cells through downregulation of poliovirus receptor (PVR/CD155), a ligand of activating NK receptor

Authors:
- Tomoko Inoue
- Katsuyuki Adachi
- Kei Kawana
- Ayumi Taguchi
- Takeshi Nagamatsu
- Asaha Fujimoto
- Kensuke Tomio
- Aki Yamashita
- Satoko Eguchi
- Haruka Nishida
- Hiroe Nakamura
- Masakazu Sato
- Mitsuyo Yoshida
- Takahide Arimoto
- Osamu Wada-Hiraike
- Katsutoshi Oda
- Yutaka Osuga
- Tomoyuki Fujii
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Affiliations: Department of Obstetrics and Gynecology, Faculty of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-8655, Japan
Published online on: July 26, 2016 https://doi.org/10.3892/ijo.2016.3631
Pages: 1297-1304
Copyright: © Inoue et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Cancer-associated fibroblasts (CAFs) play an important role in cancer expansion and progression in tumor microenvironment (TME), via both direct and indirect interactions. Natural killer (NK) cells play a crucial role in anticancer immunity. We investigated the inhibitory effects of CAFs on NK cell activity. CAFs were isolated from endometrial cancer tissue, while normal endometrial fibroblasts (NEFs) were obtained from normal endometrium with no pathological abnormality. NK cells were obtained from allogenic healthy volunteers. CAFs or NEFs were co-cultured at an NK/fibroblast ratio of 1:1 with or without inserted membrane. For NK cell activity, K562 cells were cultured as target cells. NK cell-killing activity was determined by calculating the ratio of PI-positive K562 cells in the presence of NK cells co-cultured with fibroblasts versus NK cells alone. To examine whether NK cell activity was suppressed by IDO pathway, we inhibited IDO activity using the IDO inhibitor 1-MT. We demonstrated that CAFs derived from endometrial cancer induced greater suppression of the killing activity of allogenic NK cells compared with normal endometrial fibroblasts (NEFs). The suppression of NK cell activity by CAFs was inhibited when a membrane was inserted between the CAFs and NK cells, but not by 1-MT, an inhibitor of IDO. We focused on receptor-ligand interactions between CAFs and NK cell and found that cell-surface poliovirus receptor (PVR/CD155), a ligand of activating NK receptor DNAM-1, was downregulated in the CAFs compared with NEFs. To confirm whether PVR downregulation results in the decrease of NK cell-killing activity, PVR expression in NEFs was knocked down using siRNA against PVR (PVRsi). NK cell activity was suppressed by co-culture with PVR-knockdown NEFs, to a similar extent than CAF-induced suppression. CAFs showed increased suppression of NK cell-killing activity compared with NEFs, due to decreased PVR cell surface expression, a ligand of an NK activating receptor. This study demonstrated a novel mechanism of suppression of NK cell activity by CAFs in the TME.

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