For sensitization of collected T-cells using mature primed DCs, cells collected from same donor were used.
Right panel: animal, in which CD11c+ DCs cells were depleted following administration of diphtheria toxin (DT).
To check the purity of DCs, cells were labeled with anti-CD11c (1:1000 dilution) monoclonal antibodies (BD).
The percentage and number of conventional DCs cells staining YAe+ is shown in (A ) and (B ) respectively.
To confirm generation of immature DCs, cells were examined by flow cytometry for expression of lineage and differentiation specific markers.
DC2.4, an immortalized DC cell line [18], has been well characterized and is widely used for immunologic research [19, 20].
On day 7, the nonadherent cells were removed and analyzed by FACS using DC cell surface markers.
Our present knowledge is essentially limited to studies in cultured DC cell lines with no real investigations in primary cells.
Results indicate that the presence of CD28 on DC cell surface, where speckles of CD28 was identified (Fig. 4D).
