Langerhans' cells produce type IV collagenase (MMP-9) following epicutaneous stimulation with haptens

Abstract

For initiation of the contact hypersensitivity response, epidermal Langerhans' cells (LC) migrate from the epidermis to draining nodes via afferent lymphatics by passing through the basement membrane. In this study, we examined production of matrix metalloproteinases (MMPs) in LC-enriched epidermal cells to clarify the type of enzymes involved in LC transmigration through the basement membrane. Using gelatine enzymography and immunoblotting analysis, 95,000 MW type IV collagenase (MMP-9) was found to be produced by LC-enriched epidermal cells. Analysis of the kinetics of MMP-9 expression showed that its production was induced within 6 hr after application of 2,4,6-trinitrochlorobenzene (TNCB), substantially increased between 12 hr and 24 hr, and then decreased to the normal level by 7 to 10 days. Other haptens, such as 2,4-dinitrochlorobenzene and 2,4-dinitrofluorobenzene, also induced MMP-9 expression. Fluoroescence-activated cell sorter analysis revealed that LC were one of the major cell types to express MMP-9 in response to TNCB. In addition, highly enriched LC from sensitized skin were shown to express strong gelatinolytic activity. These results indicate that LC by themselves, as well as other epidermal cells, are capable of producing MMP-9, and suggest that MMP-9 may contribute to proteolysis associated with transmigration of LC in the induction phase of contact dermatitis.