What is the best way to get fresh isolates from PBMC’s?
It is best to stimulate the PBMCs with PHA (5 mg/ml) for 48 hours and then add 10% human IL-2. Because the virus resides in the T-cells and monocytes, if the T-cells and monocytes are separated, you will have better success. If PHA stimulated cord blood cells are available, then use them to co-cultivate with PBMCs. The appearance of large, multinucleated giant cells is a good indication that these are infected cells. Monitor them with HHV-6 specific monoclonal antibodies (MABs).
