Visualization and Characterization of Migratory Langerhans Cells in Murine Skin and Lymph Nodes by Antibodies Against Langerin/CD207

Abstract

Dendritic cells are professional antigen-presenting cells that initiate primary immunity. Migration from sites of antigen uptake to lymphoid organs is crucial for the generation of immune responses. We investigated the migratory pathways specifically of epidermal Langerhans cells by tracing them from the epidermis to the draining lymph nodes. This was possible with a new monoclonal antibody, directed against murine Langerin/CD207, a type II lectin specific for Langerhans cells. In situ, resident, and activated Langerhans cells express Langerin in the epidermis and on their way through dermal lymphatic vessels. Both emigrated and trypsinization-derived Langerhans cells expressed high levels of Langerin intracellularly but reduced it upon prolonged culture periods. Sizeable numbers of Langerin+ cells were found in skin draining lymph nodes but not in mesenteric nodes. Langerin+ cells localized to the T cells areas and rarely to B cell zones. Numbers of Langerin-expressing cells increased after application of a contact sensitizer. In the steady state, Langerhans cells in the skin-draining nodes expressed maturation markers, such as 2A1 and costimulatory molecules CD86 and CD40. These molecules, CD86 and CD40, were further upregulated upon inflammatory stimuli such as contact sensitization. Thus, the novel anti-Langerin monoclonal antibody permits the unequivocal visualization of migratory Langerhans cells in the lymph nodes for the first time and thereby allows to dissect the relative immunogenic or tolerogenic contributions of Langerhans cells and other types of dendritic cells.

Langerhans cells cross-present antigen derived from skin

“…Shibaki et al (29) used mice expressing the whole OVA protein under the K14 promoter and showed that DCs emigrating from skin explants cross-presented OVA peptides. DCs emigrating from whole skin explants are a mixture of dermal DCs and LCs (30), and therefore, the relative contribution of LCs could not be determined in this report. In the second report by Mayerova et al (31), LCs were sorted from skin draining lymph nodes of mice expressing SIINFEKL under the K14 promoter.…”

IgE-Mediated Mast Cell Activation Induces Langerhans Cell Migration In Vivo

“…Henri et al (34) have demonstrated that the numbers of Langerin-positive cells in LNs draining the skin are normally regulated by a steady state of migration, consistent with the presence of some Langerin/CD207-positive cells in the control LNs. In addition, Stoitzner et al (35) have shown that Langerin/CD207-positive cells in the skin-draining LNs are mostly mature. Changes in LC numbers in both the ear epidermis and draining LNs were consistent with IgE-mediated mast cell activation inducing LC migration in the context of a background level of continuous recruitment of immature LC to the skin and subsequent migration of cells to the draining LNs.…”

Calcitonin Gene-Related Peptide Is an Important Regulator of Cutaneous Immunity: Effect on Dendritic Cell and T Cell Functions

“…Skins were cultured in 24-well plates, as described (16), in the presence of CGRP and/or MIP-3b for 72 h, and the cells that migrated into the culture medium (RPMI 1640 supplemented with 10% FCS, 0.02 mM kanamycin, and 5 mM 2-ME) were counted.…”

“…After applying a hapten, activated LCs migrated from the epidermis to the dermis and then to the draining LNs, wherein they induced T cell sensitization (16). To determine whether CGRP could affect Langerin + cell migration at the sensitization phase of CHS, we examined the effect of CGRP on the number of Langerin + cells that migrated to the draining LNs.…”