TNF-α, RANTES, and MCP-1 are major chemoattractants of murine Langerhans cells to the regional lymph nodes

We have previously reported that lymph node cells generated chemotactic factors for Langerhans cells (LCs) in the induction phase of contact dermatitis. In order to clarify the chemotactic factors involved in migration to the regional lymph nodes, we investigated the migratory activity of murine LCs toward several cytokines and chemokines in vitro. One-day cultured LC-enriched epidermal cells were added to the upper compartment of a modified Boyden chamber and cytokines were added to the lower compartment. We counted dendritic cells migrated to the lower chamber as LCs under phase contrast microscopy. About 99% of migrated dendritic cells were positively reacted with anti-Ia(d) and NLDC145 antibodies and considered to be LCs. We could detect LC migration more accurately by this direct examination than by counting the migrated cells in the filter membrane of a Boyden chamber. In our system, migration of murine epidermal LCs was stimulated by TNF-alpha, RANTES and MCP-1, but not by GM-CSF, IL-1 beta, IL-4, and IL-6. TNF-alpha induced LC migration at concentrations from 4X10(3) U/ml to 5x10(4) U/ml. RANTES at concentrations from 10 to 100 ng/ml and MCP-1 at a concentration of 100 ng/ml induced LC migration in a dose-dependent manner. These data confirmed that TNF-alpha, RANTES, and MCP-1 induced LC migration from epidermis during contact sensitization.