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Background: Successful immunotherapeutic intervention could be enhanced by understanding the immune cell component of the tumour microenvironment. The aim of this study was to characterise intra-tumoural immune cell populations in a cohort of colorectal cancer patients and correlate the frequency of key immune cell subsets with clinical/pathological findings Methods: Fresh biopsy and resection samples were removed from colorectal cancer patients (n=5) and processed to single cell suspensions following pathological inspection. Informed consent was obtained from all patients prior to sampling. Single cells were incubated with fluorochromeconjugated monoclonal antibodies against the main immune cell subset-distinguishing surface proteins; namely, CD4 (T cells), CD8 (T cells), CD56 [natural killer (NK) cells], CD11b (macrophages) and CD11c [dendritic cells (DCs)]. Frequency (%) of these immune cells was analysed by flow cytometry (using a BD FACSCanto II). Results: Immune-profiling was performed on centre and leading edge colorectal tumour tissue, with non-cancerous adjacent normal tissue serving as a control. Comparison of infiltrating immune cells between these 3 tissues in a patient with a mucinous tumour (60%) at stage T4N0 revealed comparable levels of CD11b+ macrophages (14.49–18.45%), CD11c+ DCs (<1%) and CD56+ NK cells (1.03–1.34%). CD4+ and CD8+ T cell frequencies, however, differed considerably. Centre tumour: 23.6% CD4+, 32.05% CD8+; leading edge: 12.4% CD4+, 17% CD8+ and normal: 10.27% CD4+, 3.25% CD8+. Degree of tissue differentiation also yielded insights with moderately differentiated adenocarcinomas having comparable frequencies of all 5 immune cell subsets in centre tumour samples. Conclusions: Intra-tumoural immune profiling coupled with pathological findings has potential prognostic benefit

AB066. Elucidating the circulating and tumour-specific immune populations in a cohort of colon cancer patients