Dendritic cells in the normal human tympanic membrane

Dendritic cells (DCs) are antigen-presenting cells that possess an outstanding capacity to initiate primary immune responses. They reside in the tissues in an immunologically immature state. Upon antigenic challenge in vivo or short-term culture in vitro, they undergo a maturation process and turn into mature "lymphoid DCs." Langerhans cells (LCs) of the epidermis were identified as members of this DC system. They have been demonstrated in cholesteatoma matrix and in inflamed tympanic membranes, but the normal tympanic membrane was hitherto thought to be devoid of them. To clarify this question, we removed 12 normal tympanic membranes postmortem and processed them for a sheet preparation. The epidermal layers were peeled off and immunostained with the following monoclonal antibodies: HLA-DR, OKT6/CD1a, and LAG (specific for the Birbeck granules of LCs). Two tympanic membranes were also processed for routine electron microscopy. In all epidermal sheets a dense network of DCs could be demonstrated. They showed a positive immunostaining reaction with HLA-DR, but a negative one with OKT6 and LAG. Thus, they differ in their immunohistochemical properties from typical epidermal LCs. At the ultrastructural level, DCs could also be identified, but without the typical Birbeck granules. This explains the negative reaction with the LAG antibody. These findings were extended and supported by a tissue culture examination of three normal tympanic membranes. After 3 days, typical "veiled" cells (ie, mature DCs), showing positive immunostaining with HLA-DR, could be recovered from the culture medium. In an oxidative mitogenesis assay, these cells displayed strong stimulatory capacity for resting T lymphocytes.(ABSTRACT TRUNCATED AT 250 WORDS)