Figure 4.

Differentiation of LC-like cells into moLCs was mediated through direct interaction with E-cadherin on KCs. (A) Three-day-LC-like cells were cultured with (right) or without (left) KCs, with (bottom) or without (top) Transwell® inserts. Cells were stained with anti-Langerin and anti-DC-SIGN. (B) Three-day-LC-like cells were cultured with KCs pretreated with anti-E-cadherin antibody or control isotype-matched antibody for an additional 3 days and stained with anti-Langerin and anti-DC-SIGN (two left-most panels). After confirming the expression of HLA-ABC on KCs, KCs were pretreated with anti-HLA-ABC antibody and co-cultured with 3-day-LC like cells (two right-most panels). (C) Three-day-LC-like cells were added to human E-cadherin-coated plates and incubated for 3 days. The generated cells were stained with anti-Langerin, anti-DC-SIGN, anti-E-cadherin, and anti-CD1a. Expression of Langerin and DC-SIGN (left), E-cadherin (middle), and CD1a (right) in Langerin⁺ cells is shown. (D) Langerin and DC-SIGN expressions of moLCs generated using murine E-cadherin-coated plates and incubated for 3 days. (E) Three-day-LC-like cells were treated with anti-E-cadherin-specific antibody for 30 min and washed three times with CCM to remove free antibody and stimulated with plate-coated purified human E-cadherin for an additional 3 days. Results shown are representative of four independent experiments.