Construction and Analysis of a DNA Cassette for Use in DNA-based Immunization: A Potential Application for Producing Antibodies Against Fasciola gigantica

Main Article Content

Chollanot Kaset Vithoon Viyanant Veerachai Eursitthichai

Abstract

Fasciola gigantica, the tropical liver fluke, is responsible for huge economic losses in the meat industry. Unfortunately, a protective vaccine against fascioliasis is not available. In this study we analyze the use of cathepsin L (CatL), the major secreted proteolytic enzyme of this parasite, for DNA-based immunization in mouse. A chimeric DNA cassette containing the coding sequence for the transmembrane/cytoplasmic (Tm/Cyt) domain of the mouse
lysosome-associated membrane protein (LAMP-1) and the CatL encoding sequence of F. gigantica was constructed in the mammalian expression vector pcDNA3.1. LAMP-1 was selected as a co-stimulatory for this study as its combination with parasite/virus-specific sequences had provoked strong antigen-specific immune responses in mice in previous studies. A gene gun device was used to introduce the recombinant plasmid into mice and the immune response was evaluated by ELISA, immunohistochemistry and immunoblot. The
results showed that the chimeric CatL-Tm/Cyt cassette induced a high titer of IgG1 and significant titers of IgG2a and IgG2b specific to CatL. This indicates a mixed Th1/Th2 response and demonstrates that pcDNA3.1-CatL-Tm/Cyt can be delivered as a functional DNA vaccine using immunization through a gene gun device.

Keywords

Article Details

How to Cite
Kaset, C., Viyanant, V., & Eursitthichai, V. (1). Construction and Analysis of a DNA Cassette for Use in DNA-based Immunization: A Potential Application for Producing Antibodies Against Fasciola gigantica. Science & Technology Asia, 15(3), 81-90. Retrieved from https://www.tci-thaijo.org/index.php/SciTechAsia/article/view/41287
Section
Articles