Browsing by Subject "TILs"

Cancer immunotherapy has advanced with the introduction of anti-PD-1 therapy. Anti-PD-1 therapy blocks the inhibitory immune checkpoint receptor PD-1, and allows T cells to regain their cytotoxic potentials in response to cancerous cells. Amongst the cytotoxic lymphocytes other than T cells, natural killer (NK) cells have not been fully explored regarding anti-PD-1 treatment. With this in mind, our aim was to characterize the immunophenotype of the immune cells in patients diagnosed with renal cell carcinoma (RCC), and separate their corresponding tumor infiltrating lymphocytes (TILs). Altogether, we analyzed the immunophenotype of the T and NK cells and RCC tumor cells from 13 RCC patients using flow cytometry. In addition, we tested the function of NK cells against RCC cells using various cell line models (NK-92 and RCC cell lines), and monitored the cytotoxicity in real time using a cell impedance assay. Based on the results, we suggest that patients diagnosed with RCC possess unique immune profiles depending on the individual, but also share characteristics regarding the tumor. In addition, NK cells have the potential to activate a cytotoxic response to RCC cells. By phenotyping the TILs, potential novel biomarkers for immuno-oncological (IO) therapy could be detected, which could aid in determining which patients would most likely benefit from immunological therapies such as anti-PD-1 treatment.

Tumors contain variable number of different immune cells that infiltrate the tumor microenvironment, such as tumor infiltrating lymphocytes (TILs). More research is needed to understand the functional and clinical importance of various TIL subgroups in cancer. Understanding the differences between individual cancer patients will help development of new treatment methods and discovering why only some patients respond to immunological treatments. Renal cell carcinoma (RCC) is the most common kidney cancer type with good overall survival prognosis when the tumor is surgically removed before it has metastasized. However, the prognosis of RCC is significantly decreased when the cancer has spread. The aim of this master’s thesis project was to characterize the tumor infiltrating lymphocyte populations in patient derived RCC samples. Characterization was done with flow cytometry and a custom antibody panel designed to detect various lymphocyte subpopulations. We also wanted to further study the TILs by expanding the lymphocytes from the tumor samples and test their function in an impendence-based assay against matched autologous tumor cells. Based on the flow cytometry results, the different RCC subtypes in the cohort showed some variation in TILs. Still, more research is needed to investigate these differences. We were able to culture the TILs from the RCC tumor samples, and most of them were CD4+ T cells expressing memory markers CD45RO and CCR7. Some expanded TILs expressed markers related to T cell activity and terminal differentiation. In conclusion, this thesis provided material and insights for future RCC TIL experiments as well as considerations for optimization needed in further studies.