Basic Study
Copyright ©The Author(s) 2016. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Apr 7, 2016; 22(13): 3573-3580
Published online Apr 7, 2016. doi: 10.3748/wjg.v22.i13.3573
Analysis of tumor-infiltrating gamma delta T cells in rectal cancer
Liang Rong, Ke Li, Rui Li, Hui-Min Liu, Rui Sun, Xiao-Yan Liu
Liang Rong, Ke Li, Hui-Min Liu, Rui Sun, Xiao-Yan Liu, Department of Gastroenterology, the Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, Xinjiang Uygur Autonomous Region, China
Rui Li, Department of Gastroenterology, the First Affiliated Hospital of Soochow University, Suzhou 215006, Jiangsu Province, China
Author contributions: Rong L designed the study; Li K performed the research; Liu HM contributed new reagents and analytic tools; Rong L and Sun R analyzed the data; Rong L and Li K wrote the paper; all the authors contributed to this manuscript.
Institutional review board statement: The study was revised and approved by the Fifth Affiliated Hospital of Xinjiang Medical University Institutional Review Board.
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of the First Affiliated Hospital of Soochow University (IACUC protocol number: 2013-10-13).
Conflict-of-interest statement: We declare that there are no conflicts of interest to disclose.
Data sharing statement: No additional data are available.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Rui Li, MD, Department of Gastroenterology, the First Affiliated Hospital of Soochow University, No. 188 Shizi Street, Suzhou 215006, Jiangsu Province, China. rongliangrl@yeah.net
Telephone: +86-512-65223637 Fax: +86-512-65223637
Received: November 1, 2015
Peer-review started: November 2, 2015
First decision: November 27, 2015
Revised: December 14, 2015
Accepted: January 11, 2016
Article in press: January 11, 2016
Published online: April 7, 2016
Abstract

AIM: To investigate the regulatory effect of Vδ1 T cells and the antitumor activity of Vδ2 T cells in rectal cancer.

METHODS: Peripheral blood, tumor tissues and para-carcinoma tissues from 20 rectal cancer patients were collected. Naïve CD4 T cells from the peripheral blood of rectal cancer patients were purified by negative selection using a Naive CD4+ T Cell Isolation Kit II (Miltenyi Biotec). Tumor tissues and para-carcinoma tissues were minced into small pieces and digested in a triple enzyme mixture containing collagenase type IV, hyaluronidase, and deoxyribonuclease for 2 h at room temperature. After digestion, the cells were washed twice in RPMI1640 and cultured in RPMI1640 containing 10% human serum supplemented with L-glutamine and 2-mercaptoethanol and 1000 U/mL of IL-2 for the generation of T cells. Vδ1 T cells and Vδ2 T cells from tumor tissues and para-carcinoma tissues were expanded by anti-TCR γδ antibodies. The inhibitory effects of Vδ1 T cells on naïve CD4 T cells were analyzed using the CFSE method. The cytotoxicity of Vδ2 T cells on rectal cancer lines was determined by the LDH method.

RESULTS: The percentage of Vδ1 T cells in rectal tumor tissues from rectal cancer patients was significantly increased, and positively correlated with the T stage. The percentage of Vδ2 T cells in rectal tumor tissues from rectal cancer patients was significantly decreased, and negatively correlated with the T stage. After culture for 14 d with 1 μg/mL anti-TCR γδ antibodies, the percentage of Vδ1 T cells from para-carcinoma tissues was 21.45% ± 4.64%, and the percentage of Vδ2 T cells was 38.64% ± 8.05%. After culture for 14 d, the percentage of Vδ1 T cells from rectal cancer tissues was 67.45% ± 11.75% and the percentage of Vδ2 T cells was 8.94% ± 2.85%. Tumor-infiltrating Vδ1 T cells had strong inhibitory effects, and tumor-infiltrating Vδ2 T cells showed strong cytolytic activity. The inhibitory effects of Vδ1 T cells from para-carcinoma tissues and from rectal cancer tissue were not significantly different. In addition, the cytolytic activities of Vδ2 T cells from para-carcinoma tissues and from rectal cancer tissues were not significantly different.

CONCLUSION: A percentage imbalance in Vδ1 and Vδ2 T cells in rectal cancer patients may contribute to the development of rectal cancer.

Keywords: Rectal cancer, T cells, Vδ1 T cells, Vδ2 T cells, Foxp3, Cytotoxicity

Core tip: The percentage of tumor-infiltrating Vδ1 T cells in rectal cancer patients increased when T stage increased, whereas the percentage of tumor-infiltrating Vδ2 T cells in rectal cancer patients decreased as T stage increased. Vδ1 T cells from rectal cancer tissues had strong regulatory effects, and in rectal cancer tissues the main infiltrating γδ T cells were Vδ1 T cells. Although Vδ2 T cells from rectal cancer tissues have strong cytotoxic effects, there was little infiltration of Vδ2 T cells in rectal cancer tissues. Thus, an immunosuppressant microenvironment was formed in rectal cancer tissues, which may limit antitumor immunity and allow tumors in rectal cancer patients to evade immune surveillance.