Methods for studying immuno-effector functions and antigen presenting activity of human macrophages

Macrophages are now recognized as cells that perform a variety of functions making them important cells in both the specific and non-specific immune responses. In the inflammatory response mature monocytes leave the bloodstream to enter macrophages and then migrate to the site of infection. This process is usually co-ordinated by the production of so-called chemotaxins that direct the movements of the macrophages. This can be measured in vitro by a chemotaxis assay that separates macrophages from the chemotactic fluid by a micropore filter. After incubation migrated cells are counted at the other side of the filter and indicate the chemotactic response. At the inflammatory site phagocytosis and killing of invading bacterial micro-organisms is an important feature of macrophages. We describe an assay to measure phagocytosing capacity using IgG-coated sheep red blood cells and a bacterial killing assay. In addition a tumor killing assay is briefly introduced. Finally, the specific immune response is initiated by presentation of antigen to antigen specific T cells by specialized cells such as dendritic cells that are closely related to macrophages. An enrichment procedure for macrophages and dendritic cells is outlined and antigen presentation assays are described.