FIGURE 2

Colorectal DCs can acquire and transport Ad-encoded gene to ILNs. (a) Two days after ICR administration of 2 10⁹ PFU of AdGFP, both colorectal LP and ILNs were isolated to determine GFP⁺ cells by flow cytometry (upper panel/top row: AdBHG control vector lacking GFP; bottom row/AdGFP: numbers indicate percentage of total cells recovered). LP and ILN cells were stained for MHC class II and CD11c (lower panel/top tow; numbers indicate percentage of total regardless of GFP⁺ or GFP^-). Gated on the GFP⁺ cell population (R1), cells double positive for MHC class II and CD11c were evaluated for gene expression (lower panel/bottom row; numbers indicate percentage of GFP⁺ cells). (b) CFSE-labeled, BM-derived DCs at various doses were injected into the colorectal Mucosa, and ILN cells were recovered 5 days later and pooled from two mice. CFSE⁺ cells were analyzed by flow cytometry. (c) BM-derived DCs were transfected with AdOVA overnight and injected into the colorectal mucosa. Five days after immunization, ILN cells were cultured without peptide stimulation for 3 days. Specific killings of ILN effector cells on EL-4/SIINFEKL target cells at indicated ratios (E:T) were assayed by Cr release assay. The results are representative of at least two independent experiments. Ad, adenovirus; DC, dendritic cell; ILN, iliac lymph node; AdGFP, Ad expressing green fluorescent protein; CFSE, carboxyfluorescein diacetate succinidyl ester; LP, lamina propria; PFU, plaque-forming unit; MHC, major histocompatibility complex; BM, bone marrow; AdOVA, Ad expressing ovalbumin.