Development of dual reporter imaging system for to monitor the spatio-temporal pathogenesis and vaccine efficacy.

Purpose: Study on the pathogen and the pathogen-related disease require the information at both cellular and organism level. However, lack of appropriate high-quality antibodies and the difference between the experimental animal models make it difficult to analyze mechanism of pathogen-related diseases. For more reliable research on the infection and immune-response of pathogen-related diseases, accurate analysis is essential to provide spatiotemporal information of pathogens and immune activity to avoid false-positive or mis-interpretations. In this regards, we have developed a method for tracking in the animal model without using the specific antibodies for the .

Materials And Methods: A dual reporter plasmid using GFP-Lux with putative bacterioferritin promoter (pBfr) was constructed and transformed to live vaccine strain to generate LVS (LVS)-GFP-Lux for both fluorescence and bioluminescence imaging. For vaccination to infection, LVS and lipopolysaccharide (LPS) from LVS were used.

Results: We visualized the bacterial replication of in the cells using fluorescence and bioluminescence imaging, and traced the spatio-temporal process of pathogenesis in mice. Vaccination with LPS purified from LVS greatly reduced the bacterial replication of LVS in animal model, and the effect of vaccination was also successfully monitored with imaging.

Conclusion: We successfully established dual reporter labeled for cellular and whole body imaging. Our simple and integrated imaging analysis system would provide useful information for analysis of infection as well as experiments, which have not been fully explained yet with various technical problems.