Photochemical Treatment of Drosophila APCs Can Eliminate Associated Viruses and Maintain the APC Function for Generating Antigen-Specific CTLs Ex Vivo

Figure 4

UVADEX-treated Drosophila cells retain APC function. (a) CD8 T cells retained CD69 expression when activated by UVADEX-treated Drosophila APCs. Purified CD8 T cells from 2C transgenic mice were cultured with APCs (Fly/L^d/B7.1/ICAM) at 37°C for 4 h. The cultured cells were collected and stained with anti-CD69-FITC mAb at 4°C and isotype control antibody for 30 minutes and analyzed for CD69 expression by FACS analysis. Left panel: CD8 T cells were stimulated by nontreated APCs. Right panel: CD8 T cells were stimulated by UVADEX-treated APCs. The number in the panel indicated the mean fluorescence intensity by FACS. (b) UVADEX-treated APCs stimulate the proliferation of CD8 T cells as efficiently as untreated APCs. CFSE-labeled CD8 T cells purified from 2C transgenic mice were cultured with APCs in the presence of 10 μM of QL9 peptide at 37°C for 2 days. The cultured cells were collected and stained with PE-conjugated anti-mouse CD8 mAb at 4°C for 30 min. CD8-positive cells were gated from FACS and further analyzed for their green fluorescence intensity. (c) CD8 T cells stimulated with UVADEX-treated APCs retain their cytolytic function. CD8 T cells purified from 2C mice were stimulated as described in Figure 1(c). 2C CD8 T cells were used in ⁵¹Cr release assay with RMAS^Ld target cells.

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