Preparation of reporter cell line for validation of recombinant protein biological activity

Natural killer cells belong to immune cells mediating non-specific immune response. On their surface, natural killer cells express many ligand-specific activation or inhibition receptors, among which belongs natural cytotoxicity activation receptor NKp30, which is able to trigger cytotoxic immune response upon interaction with its ligands. Natural cytotoxicity receptor NKp30 comprises of one immunoglobulin-like ligand-binding extracellular domain with short stalk domain, allowing the extracellular domain of NKp30 receptor to form oligomers, which leads to strengthening of ligand-receptor interaction. Furthermore, signalization leading to cell activation is affected by oligomerization of the extracellular domain of NKp30 receptor. This thesis focuses on preparation of constructs inspired by chimeric antigen receptor theory that carry extracellular domain of NKp30 receptor with stalk domain or only with ligand- binding domain and on validation of biological activity of prepared constructs after expression on the cell membrane. Two vectors for expression of the constructs inspired by chimeric antigen receptors carrying genes for extracellular domain of NKp30 receptor were prepared and to validate their biological activity, they were transfected into Jurkat cells. Key words: NK cells, NKp30, reporter..