Study On Genetically Engineered Subunit Vaccines Against Swine Vesicular Disease And Classical Swine Fever

Both the Swine vesicular disease (SVD) and Classical swine fever (CSF) are two list A diseases defined by Office International des Epizooties (OIE) due to their high infectivities to pigs and damages to pig-feeding industry. SVD is another highly contagious disease of pigs, which its cilinical symptom is similar to the one causes by Foot and mouth disease (FMD) so that it has been considering as economically important during pig products currency and international trade. CSF is an acute or chronic disease, which usually causes fever and haemorrhages of infected pigs. A well-known lapinized Chinese strain of CSF vaccine, which also called C strain vaccine or C vaccine, has been proved to be highly efficient protecting pigs from Classical swine disease, whereas the shortcoming is that the vaccinated animals can`t be distinguished from wild virus infected ones because the antibody induced by C-vaccine is similar to the recovered animals from wild virus infection ever.So far there is no available viaccine agansit SVD reported yet, therefore developing an efficient, safe and label genetical engineer vaccines and corresponding diagnosis tools for both diseases has become the key problems to be solved.In this study, genetical engineer subunit vaccines of SVD and CSF were developed by employing molecular biological tools, and their immunogenicity has been primarily evaluated by immunizing experimental animals. The main research process as following:1.Recombinant plasmid pBABE puro-P1 were constructed, following artificial retroviral particles were obtained by co-transfection recombinant plasmid pBABE puro-P1 and pVSV-G envelope vector into the GP2-293 package cells. PK-15 cells lines were transduced by virus mediated by polybrene(8μg/mL), and then positive cell clones were selected by puromycine selecting approach. P1 protein expressed in PK-15 cells can be recognized by SVD positive serum has been proved by indirect immunofluorescence assay, nevertheless P1 gen can be amplified from different generation (1st,8th,16th and 30th) positive cell genome by PCR, which indicates that positive cells could stably carry target genes in subsequent passaging.2.Plenty of harvested cell culture were emulsificated by Freund`s adjuvant to immunize guinea pigs, and efficiency of vaccines were evaluated by lymphocytes proliferation assay,...