Strategies and Techniques for NK Cell Phenotyping

Therapies based on activating the immune system, that is, immunotherapy, are now widely implemented in clinical praxis in patients with advanced cancer. Although cancer immunotherapy can result in long-lasting clinical responses, the majority of patients do not respond or develop resistance. Furthermore, cancer immunotherapy is being increasingly combined with other forms of immunotherapy or conventional cancer therapies. It is therefore much needed to identify biomarkers that can precisely classify what patients will benefit from the treatment without any major adverse events and to further develop the efficacy of cancer immunotherapy. While much attention has been focused on monitoring T cell responses in cancer immunotherapy, recent reports have shown that NK cells also play a major role in the response to cancer immunotherapy. The gold standard for immunoprofiling of NK cells is flow cytometry, but other technologies have emerged and include mass cytometry, multiplex immunohistochemistry, and single-cell RNA-sequencing. In this chapter we provide a detailed protocol to profile NK cells using flow cytometry, and a brief introduction to other techniques.