Protocols for Innate Lymphoid Cell Phenotypic and Functional Characterization: An Overview

The innate immune system forms a first line of defense against infections and is an essential component in immune defense prior to the generation of sustained adaptive immune responses. Until recently this innate response was believed to be predominantly elicited by cells of myeloid origin. The last 10 years have seen the discovery of an extensive family of innate lymphoid cells (ILCs) characterized by the ability to express cytokines, traditionally associated with activated T effector cells, and lack of expression of antigen receptors.ILCs are enriched at barrier surfaces and rapidly respond to alarmins within the tissue microenvironment. ILCs lack recombinant activating gene (RAG)-dependent rearranged receptors, lack myeloid and dendritic cell lineage defining factors, and possess lymphoid morphology. The first characterized innate lymphoid cells were natural killer cells (NK) cells and lymphoid tissue inducer (LTi) cells. NK cells, defined in 1975), mediate important early antigen independent immune responses. By contrast, LTi cells, defined in 1997, are vital for formation of lymph nodes during embryogenesis. In the past decade, research on ILCs has defined them as important regulators of barrier immunity. However, ILC phenotype and functional characterization is complex and requires highly specific protocols for delineating ILC specific function from other well-characterized immune cells. Within this book, each chapter provides an in-depth protocol that will expand on techniques used by laboratories to study ILC development, characterization and function in mice and humans.