Histologic analysis of T lymphocyte activation in reactive lymph nodes

As an approach to defining the anatomic sites of T cell activation in situ, we have developed an immunocytochemical stain for IL-2, a T cell-derived cytokine synthesized shortly after Ag-induced activation. Analysis of lymph nodes from mice immunized with keyhole limpet hemocyanin emulsified in CFA demonstrates that the IL-2⁺ cells appear in a perivascular location 4 days after antigenic challenge. After germinal centers develop, IL-2⁺ cells are situated in a parafollicular pattern. Serial sections stained for different types of APC, including B cells, interdigitating dendritic cells, and macrophages, demonstrate a close physical association between IL-2⁺ cells and macrophages. These findings may have important implications for defining how APC bearing processed Ag and Ag-specific T cells interact in the complex environment of lymphoid tissues.