We hypothesize that T cell mediated tumor clearance is associated with receptor aggregation and activation of T cells in small regions surrounding a growing tumor mass, indicating a local site of effective antigen presentation. The formation of such a pseudo-lymphoid compartment may act as a staging area at which effector T cells are re-stimulated and at which innate immune cells aggregate. We suggest that understanding of the fundamentals of cellular and molecular dynamics at the border of two model tumors will provide insight into the nature of effector tumor surveillance and rejection.
1. To define the dynamics of T cell, antigen-presenting cell and macrophage migration in or around a developing melanoma tumor mass. The goal is to define the dynamics of T cell or antigen-presenting cell migration in the tumor microenvironment using real-time imaging techniques.
2. To compare the dynamics of cells at the border before and after immunotherapies, which promote tumor rejection. The goal is to compare T cell dynamics before and after treatments or conditions that favor the tumor or the immune response and to determine whether T cells exhibit prolonged encounters, increased swarming or more tumor penetration during a productive response.
These studies will provide a high-resolution spatio-temporal map of cellular localization, migration, and interaction. These aims will provide a more complete insight into an immune recognition event than was previously possible and will help direct existing and novel therapeutic regimens.
1. Tumor Models
2. 2-Photon 4D Imaging
3. CFP (Blue) Labeling of Tumor Targets
4. Labeling of T cells and their key signaling molecules
5. Labeling of Antigen Presenting Cell: Labeling of Activated APCs with IL-12 p40-YFP mice, Mac-1 GFP mice, or Fluorescent Dextran