PCR analysis for provirus in DCs and T lymphocytes. (A) Analysis of HIV provirus load by limiting-dilution nested PCR on purified T lymphocytes, myDCs, and pcDCs. Results from 3 patients of 14 tested are shown. Lanes 1, 7, and 13 were amplified with 5000 cell equivalents of DNA; lanes 2, 8, and 14 were amplified with 1000 cell equivalents of DNA; lanes 3, 9, and 15 were amplified with 200 cell equivalents of DNA; lanes 4, 10, and 16 were amplified with 40 cell equivalents of DNA; lanes 5, 11, and 17 were amplified with 8 cell equivalents of DNA; and lanes 6, 12, and 18 were amplified with 1.6 cell equivalents of DNA. (B) DNA preparations from myDCs of 2 patients, (patient 1, lanes 1, 2, 5, 6; patient 2, lanes 3, 4, 7, 8) analyzed for integrated provirus. Lanes 1 through 4 employed Alu primers in the first-round reaction. In lanes 5 through 8, Alu primers were omitted from first-round reaction. Lanes 1, 3, 5, and 7 were amplified with 5000 cell equivalents of DNA. Lanes 2, 4, 6, and 8 were amplified with 1000 cell equivalents of DNA. Lane 9 is an unnested PCR product obtained using the second-round LTR primers and DNA extracted from H9 cells infected with HIV-1 IIIB.