6th International AIDS Conference San Francisco, California, USA — June 20-23, 1990

Int Conf AIDS 1990 Jun 20-23; 6:320 (abstract no. 1025)
Hassan HJ, Chelucci C, Leonardi A, Gringeri A, Mannucci PM, Peschle C; Dept. of Hematology-Oncology, Istituto Superiore di Sanita, Rome, Italy

OBJECTIVE: We have studied two matched groups of hemophilia A patients, comprising 25 seronegative and 18 seropositive subjects: all were similarly treated in '81-'85 with commercial not virus-inactivated factor VIII concentrates (greater than 75% equivalent batches were used in all patients). Since the possibility has been raised that high-risk seronegative individuals may carry proviral HIV-1 sequences in a dormant stage, we used the polymerase chain reaction (PCR) to detect HIV-1 DNA in these hemophilia groups.

METHODS: Serological assays were performed by ELISA and Western blotting. Isolated lymphocytes from seronegative samples were stimulated with PHA and activated with IL-2. 2-5 ug of DNA were subjected to PCR with oligo primers for the conserved "gag" and "env" regions. The amplified regions were separated by 1.5% agarose gel electrophoresis. DNA was hybridized with an oligo internal to the amplified regions.

RESULTS AND CONCLUSIONS: No hybridizations signal was detected in all seronegative samples after two-fold 30-cycles amplification in both monocytes and lymphocytes, as well as in stimulated T-cells. In all seropositive samples a clear hybridization signal was observed after the first 30 cycles, although with different intensities (a substantially equivalent signal with the same samples were observed using primers specific for the factor X gene): these differences may reflect different numbers of infected cells and/or proviral sequences in a single cell.

900620
1025

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