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

Figure 2

UVADEX treatment inactivates insect and Drosophila viruses. (a) Baculovirus-infected Sf9 cells were treated with UVADEX for 5 min or left untreated and grown for 4 days. Supernatant from infected cells was collected and used to reinfect Sf9 cells in a 96-well plate. Virus was detected by gp64 antibody staining using a rapid microtiter assay kit (BD BacPAKTM Baculovirus Rapid Titer Kit). Infected cells were pictured under a microscope. Left panel: viral infected cells. Right panel: UVADEX pretreated viral infected cells. (b) Inactivation of virus in Drosophila virus X-infected clone 5-5 cells by UVADEX. Drosophila viral-free cell line clone 5-5 was infected with UVADEX-pretreated or non-treated Drosophila viral fraction at RT for 1 h. The infected cells were washed with PBS completely to remove any residual viral particles and then cultured in media for 3 days. Infected cells were pictured under a microscope. Left panel: viral infected non-UVADEX-treated cells. Right panel: UVADEX-pretreated viral infected cells. (c) PI staining of UVADEX-pretreated or UVADEX-nontreated Drosophila viral fraction-infected clone 5-5 cells at day 3. (d) Drosophila lytic viral-free cell line clone 5-5 was infected with different dilutions of Drosophila viral fraction (1 : 1 dilution from stock as indicated to 2⁻¹) pretreated with 5 μg/ml of UVADEX plus UV for 10 mins. Infected cells were cultured for 3 days. Cells were collected and stained with PI (1 μg/1 × 10⁶ cells) at 4°C for 10 min to determine the percentage of cell survival cells.

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