title: Analysis of tumor-infiltrating gamma delta T cells in rectal cancer.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  A percentage imbalance in Vδ1 and Vδ2 T cells in rectal cancer patients may contribute to the development of rectal cancer.
authors.affiliation: 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.
authors.forename: Liang
authors.initials: L
authors.lastname: Rong
authors.affiliation: 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.
authors.forename: Ke
authors.initials: K
authors.lastname: Li
authors.affiliation: 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.
authors.forename: Rui
authors.initials: R
authors.lastname: Li
authors.affiliation: 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.
authors.forename: Hui-Min
authors.initials: HM
authors.lastname: Liu
authors.affiliation: 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.
authors.forename: Rui
authors.initials: R
authors.lastname: Sun
authors.affiliation: 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.
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Liu
mesh_terms: D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D018920:Coculture Techniques; D003602:Cytotoxicity, Immunologic; D006801:Humans; D008213:Lymphocyte Activation; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology* / Q000378:metabolism; D009367:Neoplasm Staging; D010641:Phenotype; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000276:immunology* / Q000378:metabolism; D012004:Rectal Neoplasms / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D013997:Time Factors
chemical_list: D016692:Receptors, Antigen, T-Cell, gamma-delta
keywords: Cytotoxicity; Foxp3; Rectal cancer; T cells; Vδ1 T cells; Vδ2 T cells