T Cell Phenotypes Associated with Oesophagogastric Adenocarcinoma

  • Carla Rengifo

Student thesis: Doctor of Medicine


T-Cell Phenotypes Associated with Oesophagogastric Adenocarcinoma Introduction: In the UK in 2016, approximately 16,000 new cases of oesophagogastric cancer were diagnosed. Chemotherapy and surgery are the mainstay of treatment but are associated with considerable morbidity and overall survival rates are dismally low. Worryingly the incidence of oesophageal adenocarcinoma is predicted to rise in the UK. Promising results using immunotherapies, including adoptive cell therapy and immune checkpoint inhibition, have been observed in other solid malignancies such as melanoma, and hold potential promise for oesophagogastric cancers. The tumour microenvironment and detectable changes in the peripheral blood of patients with oesophagogastric adenocarcinoma have not been widely researched in a Western population, potentially due to prevalence being higher in Eastern countries such as Japan and China. A better understanding of the tumour immune relationship in oesophagogastric adenocarcinoma may lead to the identification of potential biomarkers and development of novel treatment strategies. Aims Based on previous research (including the host laboratory), the primary aim was to quantify levels of regulatory T-cells, T helper 17 and 22 cells; relate expression to prognosis; and explore as potential predictive biomarkers. The secondary aim was to assess the feasibility of expanding tumour infiltrating lymphocytes (TIL) and perform in vitro experiments to evidence the potential for ACT in the treatment of oesophagogastric adenocarcinoma. Methods: T-cell phenotype markers levels were determined using multi-colour flow cytometry in the peripheral blood of patients with oesophagogastric cancer before and after initiation of chemotherapy, which was compared to healthy volunteers. Experiments to isolate and expand TIL from tumour specimens were performed using the GentleMACs system of dissociation. Results: Chapter 3 (Results 1) characterised 100 patients with oesophagogastric adenocarcinoma who were enrolled for the peripheral blood phenotyping arm of this thesis and reported poor survival patterns confirmatory with the literature. Chapter 4 (Results 2) reported that higher levels of Th17, Th22 and TCRgd cells were demonstrated in the peripheral T-cells of patients with cancer compared to healthy controls. Th17 levels appear to be predictive of survival outcome at 21-months for patients with oesophageal adenocarcinoma. Chapter 5 (Results 3) reported higher levels of Treg, CD4+OX40+, TregICOS+ and CD8+ICOS+ cells in analysis of the same patient cohort; however, levels were unrelated to site or stage. A decrease in Treg and Th22 levels were also observed in patients after initiation of chemotherapy. Chapter 6 (Results 4) showed that cytotoxic TILs were successfully extracted and expanded from three of thirteen tumour specimens. Th17 levels were significantly increased in these expanded samples. Conclusion: This thesis has performed complex peripheral T-cell phenotyping in potentially the largest group of patients with oesophageal adenocarcinoma from the UK, this is unique. This research demonstrated interesting differences in the immune phenotype of circulating PBMCs in patients with oesophagogastric adenocarcinoma using simultaneous expression of multiple markers included in Panel A and B. Th17 levels have been identified as a marker which correlates with clinical outcome and this finding should be expanded in further studies. Although this study identified problems in extracting and expanding TIL, it demonstrated that the technique is possible and therefore future work should optimise this protocol further to evidence the potential for ACT in oesophagogastric adenocarcinoma. By continuing to monitor the changes in the immune system and research into the potential role of adoptive cell therapy in this disease, potentially novel immunotherapies will change the prognosis of what is an extremely deadly disease.
Date of Award31 Dec 2019
Original languageEnglish
Awarding Institution
  • The University of Manchester
SupervisorAndrew Renehan (Supervisor), Anne Armstrong (Supervisor) & Gray Kueberuwa (Supervisor)


  • Tumour infiltrating lymphocytes
  • Multi colour flow cytometry
  • Adoptive Cell Therapy
  • Oesophageal adenocarcinoma
  • Oncoimmunotherapies
  • T-cell
  • Gastric adenocarcinoma

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