METHODS FOR STUDYING IMMUNE-EFFECTOR FUNCTIONS AND ANTIGEN-PRESENTINGACTIVITY OF HUMAN MACROPHAGES

Macrophages are now recognized as cells that perform a variety of func tions making them important cells in both the specific and non-specifi c immune responses. In the inflammatory response mature monocytes leav e 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. Th is can be measured in vitro by a chemotaxis assay that separates macro phages from the chemotactic fluid by a micropore filter. After incubat ion migrated cells are counted at the other side of the filter and ind icate the chemotactic response. At the inflammatory site phagocytosis and killing of invading bacterial micro-organisms is an important feat ure of macrophages. We describe an assay to measure phagocytosing capa city using IgG-coated sheep red blood cells and a bacterial killing as say. 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 cel ls that are closely related to macrophages. An enrichment procedure fo r macrophages and dendritic cells is outlined and antigen presentation assays are described.