Study On The Construction, Immunogenicity And Safety Of Taenia Solium Oncosphere Gene Vaccine Vector Of TSOL18

Cysticercosis is an important zoonosis caused by Taenia solium larval cysticercus cellulosae that can infect intermediate hosts such as pigs or human, which is a major public health problem in most areas of China, and a great potential threat to human security. This disease, whose prevention is always an obstacle for veterinarians mainly because of limitation of protective antigens, is known as an economic disease for it attributes not only to great economic loses but also to decrease of international competition on meat products.1.In this study, recombinant expression plasmid pVAX1/TSOL18 was constructed and then transfected into BHK-21 cell line with liposome 2000. The results of SDS-PAGE, Western blotting and fluorescence antibody assay showed that TSOL18 protein was successfully expressed in BHK-21 cell line and could be recognized by positive serum against cysticercosis.Immunization in mice indicated that the expression vectors could efficiently induce the specific cellular and humoral immunoresponse, which laid a solid foundation for the further development of new vaccines of cysticercosis.2.The biosafety and tissue distribution of recombinant expression plasmid was discussed as a DNA vaccine in mouse. Which is one of the key questions which should be solved before it is to be put into practice in the clinical trail. In this test, we investigated the recombinant TSOL18 plasmid in order to evaluate its distribution and two main aspects of biosafety in mice. Recombinant pVAX1/TSOL18 plasmid was injected through the intramuscular introduction, then the mice were slaughtered to extract the gDNA of all kinds of tissues at the different time. The distribution and the possibility of integration of recombinant pVAX1/TSOL18 plasmids in the mice genome were analyzed by PCR technology. One day and five days after immuning the recombinants . the interesting gene of plasmid was amplified from the different gDNA of mice tissues. But different individuals provided the end fragment with different tissue. In the blood, we could amplify the end fragment all the time. Contrast to the first day, the concentration of fragment are darker in fifth day. Fifteen days later, only in one mice we could amplify the fragment of plasmid TSOL18. Thirty days later, we could detect no signals of recombinant plasmid in all the mouse. And we never detected the TSOL18 plasmid at first and fifth day in the feces. Recombinant plasmid pVAX1/TSOL18 is safe to both mice and the surrounding environment as a vaccine.