Fig. 8.

Analysis of vector persistence and immune response to vectors. a The HAdV5-PUMA and BV^CAR DNA genomes in synovial tissues were extracted from synovial tissues samples of rats killed at day 21 after intra-articular administration of HAdV5-PUMA alone (B2, B4, B5 and B11) or BV^CARHAdV5-PUMA (B10, B12, B13 and B14), and used for PCR analysis to detect the viral genomes. a The 465-bp amplified fragment corresponding to the HAdV5 hexon gene (a, arrowhead) was found in all the samples and in the positive control. Note the occurrence of a non-specific amplified fragment of 1500 bp (black dots). b, For the BV^CAR genome, no specific band of 1.1 kbp, corresponding to the baculovirus CAR gene was detected in any of the samples, but was present in the positive control. b To evaluate the immune response of the treated animals to BV^CAR and HAdV5-PUMA vectors, the sera of the animals treated for 4 days and 21 days with HAdV5-PUMA alone or with BV^CAR HAdV5-PUMA were analysed by ELISA for the presence of antibodies against adenovirus and baculovirus. A single serum dilution (1:10) was used for all samples. No significant adenovirus or baculovirus antibodies were detected in the sera of the animals treated for 4 days (grey bars, c,d). In the animal group treated for 21-days, adenovirus antibodies were detected in the sera of 5/8 animals (black bars, c) and baculovirus antibodies detected in the sera of 2/8 animals (black bars, d). Similar results were obtained in an independent experiment