FIGURE 3.

Expression of hCLEC9A on BDCA3⁺ DC and a subset of CD14⁺CD16^- monocytes. A, RT-PCR analysis showing expression of CLEC9A in most tissues, but predominantly in the brain, spleen, and thymus. B, using a novel monoclonal antibody (9A11), CLEC9A was found to be expressed on a minor population of PBMC (see supplemental Fig. S4C), and cells expressing this receptor were therefore sorted prior to analysis by multicolor flow cytometry. Using various markers, as indicated, all CLEC9A⁺ cells (gated) were found to be CD4⁺HLA-DR⁺CD11c⁺, and further analysis indicated that the receptor was predominantly expressed on BDCA3⁺ dendritic cells (59 ± 10%), but also on a subset of CD64⁺CD11b⁺CD14⁺CD16^- monocytes (14 ± 6%) and a third population of CD64⁺CD11b⁺ CD14^- CD16^- cells (23 ± 3%), which were not identified further. The data shown are representative of data obtained from at least six different donors.