Approximately 2610 5 pDCs were exposed to rotavirus particles or 4 mg/mL CpG ODN 2395 (Coley Pharmaceuticals, Ottawa, Canada), as indicated, for 1 hour in serum-free RPMI-1640 (CellGro, Manassas, VA) supplemented with penicillin/streptomycin and L-glutamine (Gibco, Carlsbad CA). pDCs were washed and suspended at a concentration of 1 million pDCs/mL in RPMI-1640 with 10% heat-inactivated fetal bovine serum (Hyclone, Logan, UT), penicillin/streptomycin, L-glutamine and 10 ng/mL IL3 (Peprotech, Rocky Hill, NJ) until harvest. Where indicated, rotavirus infection was blocked using neutralizing monoclonal antibodies to VP5 (mAb 2G4) or VP7 (mAb 159), or as a non-neutralizing control, a VP6 monoclonal antibody (mAb 1E11). In studies determining whether pDC infection was productive, highly-purified pDCs (mean purity: 91.51%) were exposed to RRV at an moi of 10 for one hour; neutralizing mAb 159 was then added for 30 minutes. pDCs were washed 3 times, and were frozen immediately or after overnight culture. The resulting pDC pellets and supernatants were freeze/thawed three times, and virus titer determined by FFU assay, as previously described [89] .