Tissue distribution of a plasmid DNA encoding Hsp65 gene is dependent on the dose administered through intramuscular delivery

In order to assess a new strategy of DNA vaccine for a more complete understanding of its action in immune response, it is important to determine the in vivo biodistribution fate and antigen expression. In previous studies, our group focused on the prophylactic and therapeutic use of a plasmid DNA encoding the Mycobacterium leprae 65-kDa heat shock protein (Hsp65) and achieved an efficient immune response induction as well as protection against virulent M. tuberculosis challenge. In the present study, we examined in vivo tissue distribution of naked DNA-Hsp65 vaccine, the Hsp65 message, genome integration and methylation status of plasmid DNA. The DNA-Hsp65 was detectable in several tissue types, indicating that DNA-Hsp65 disseminates widely throughout the body. The biodistribution was dose-dependent. In contrast, RT-PCR detected the Hsp65 message for at least 15 days in muscle or liver tissue from immunized mice. We also analyzed the methylation status and integration of the injected plasmid DNA into the host cellular genome. The bacterial methylation pattern persisted for at least 6 months, indicating that the plasmid DNA-Hsp65 does not replicate in mammalian tissue, and Southern blot analysis showed that plasmid DNA was not integrated. These results have important implications for the use of DNA-Hsp65 vaccine in a clinical setting and open new perspectives for DNA vaccines and new considerations about the inoculation site and delivery system.     

The impact of intralacustrine introductions with regard to space utilization and competition for territories to a cichlid fish community in Lake Malawi,Africa

At Thumbi Island West in Lake Malawi, a number of cichlid species was introduced from elsewhere in the lake some 20 years ago. These are now, in part, well established and could have an effect on the indigenous fish fauna. Repeated removals of the males of the introduced species from their territories were undertaken and artificial miniature reefs were established to find out whether the emptied existing territories or the newly established potential territories would be occupied by indigenous species. The results show that the emptied territories were almost invariably re-occupied by either a male of the introduced species conspecific to the previous territory holder or they remained empty. The newly established miniature reefs were not occupied by territorial males of either introduced or indigenous species but by young fish of a variety of species.     

Homing and home-site fidelity in rock-dwelling cichlids (Pisces: Teleostei) of Lake Malawi,Africa

Synopsis Existence of home site fidelity and homing ability was established for Pseudotropheus aurora. Territorial males stayed up to 1 1/2 years within the same territories. Females showed long-term fidelity to certain places. Transplantation experiments revealed that P. aurora males home over distances of up to at least 2500 m but would not cross deep water. The territorial males of five other sympartic species returned from 1000 m to their territories. The mechanisms and functions of homing in cichlids are discussed.