Lack of KSHV reactivation during treatment-induced immunosuppression in MM patients. (A) DNA samples extracted from BM before and 90, 180, and 360 days after second HDC were amplified by PCR using the KSHV 330233 primers. Each PCR was performed on 1 μg of genomic DNA and the amplification products were transferred to a nylon membrane and hybridized with a 32P end-labeled internal probe. The positive control (lane C) was the PCR product from the KSHV-infected BCBL-1 cell line. (B) Lanes 1 through 5 contain 10-fold dilutions of BCBL-1 DNA from 1 ng (lane 1) to 0.1 pg (lane 5). BCBL-1 DNA was diluted in the DNA extracted from heparinized BM mononuclear cells from patient no. 1 90 days after the second HDC, so that all PCR were run on 1 μg of total DNA. These data are representative of 10 experiments performed with the heparinized DNA samples extracted from the 10 patients 90 days after second HDC.
